Human PAC1 antibodies

ABSTRACT

Antibodies and antigen-binding fragments thereof that bind to human PAC1 are provided. Nucleic acids encoding the antibodies and antigen-binding fragments thereof, vectors, and cells encoding the same are also provided. The antibodies and antigen-binding fragments thereof can inhibit binding of PAC1 to PACAP, and are useful in a number of PAC1 related disorders, including the treatment and/or prevention of headache disorders, including migraine.

PRIORITY

The present application is a divisional of U.S. application Ser. No.14/304,559, filed Jun. 13, 2014, now pending, which is a continuation ofU.S. application Ser. No. 14/203,317, filed Mar. 10, 2014, nowabandoned, which claims priority to U.S. Provisional Application No.61/792,678, filed Mar. 15, 2013, all of which are hereby incorporated byreference in their entireties.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted electronically in ASCII format and is hereby incorporated byreference in its entirety. Said ASCII copy, created on May 17, 2016, isnamed A-1804-US-DIV_ST25.txt and is 242,619 bytes in size.

BACKGROUND

There is a significant unmet need for effective therapies, particularlyprophylaxis therapies, for migraine. Results from multiple studiesindicate that ˜14 million migraineurs in the US could qualify andbenefit from an effective and safe preventive therapy. Currentlyapproved migraine prophylactic therapies are only partially effectiveand have considerable side-effect profiles which significantly limit theacceptability of these medications. Despite these limitations, 4.5million individuals with frequent migraine headache in the US takeprophylactic medication for their migraines.

Pituitary adenylate cyclase-activating polypeptides (PACAP) are 38-aminoacid (PACAP38), or 27-amino acid (PACAP27) peptides that were firstisolated from an ovine hypothalamic extract on the basis of theirability to stimulate cAMP formation in anterior pituitary cells (MiyataA, Arimura A, Dahl R R, et al. Isolation of a novel 38residue-hypothalamic polypeptide which stimulates adenylate cyclase inpituitary cells. Biochem Biophys Res Commun. 1989; 164:567-574; MiyataA, Jiang L, Dahl R D, et al. Isolation of a neuropeptide correspondingto the N-terminal 27 residues of the pituitary adenylate cyclaseactivating polypeptide with 38 residues (PACAP38). Biochem Biophys ResCommun. 1990; 170:643-648). PACAP peptides belong to the vasoactiveintestinal polypeptide VIP-secretin-hormone-releasing hormone(GHRH)-glucagon superfamily with the sequence of human PACAP27 shares68% identity with vasoactive intestinal polypeptide (VIP) (Campbell R Mand Scanes C G. Evolution of the growth hormone-releasing factor (GRF)family of peptides. Growth Regul. 1992; 2:175-191). The major form ofPACAP peptide in the human body is PACAP38 and the pharmacology ofPACAP38 and PACAP27 has not been shown to be different from each other.Unless indicated otherwise herein, PACAP or PACAP38 will be used torepresent PACAP38, and PACAP27 will be used to specify PACAP27.

Three PACAP receptors have been reported: one that binds PACAP with highaffinity and has a much lower affinity for VIP (PAC1 receptor, or simply“PAC1”), and two that recognize PACAP and VIP essentially equally well(VPAP1 and VPAC2 receptors, or simply “VPAP1” or “VPAC2”, respectively)(Vaudry D, Falluel-Morel A, Bourgault S, et al. Pituitary adenylatecyclase-activating polypeptide and its receptors: 20 years after thediscovery. Pharmacol Rev. 2009; September 61(3):283-357).

As can be appreciated from the data in the table below, PACAP is capableof binding all three receptors with similar potency and is thus notparticularly selective. VIP, on the other hand, binds with significantlyhigher affinity to VPAC1 and VPAC2, as compared with PAC1. In additionto endogenous agonists PACAP and VIP, maxadilan, a 65 amino acid peptideoriginally isolated from the sand-fly, is exquisitely selective for PAC1compared with VPAC1 or VPAC2.

PACAP Receptors & Agonists

Receptor PAC1 EC50 VPAC1 EC50 VPAC2 EC50 agonist (nM) (nM) (nM) PACAP0.03 0.03 0.06 VIP 2.3 0.02 0.08 Maxadilan 0.06 >1000 >1000

SUMMARY

In one aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising: (A) a light chain CDR1 comprising (i) an amino acid sequenceselected from the group consisting of the LC CDR1 sequences set forth inTable 5A, (ii) an amino acid sequence at least 90% identical to an aminoacid sequence selected from the group consisting of the LC CDR1sequences set forth in Table 5A, or (iii) an amino acid sequence atleast 95% identical to an amino acid sequence selected from the groupconsisting of the LC CDR1 sequences set forth in Table 5A; (B) a lightchain CDR2 comprising (i) an amino acid sequence selected from the groupconsisting of the LC CDR2 sequences set forth in Table 5A, (ii) an aminoacid sequence at least 90% identical to an amino acid sequence selectedfrom the group consisting of the LC CDR2 sequences set forth in Table5A, or (iii) an amino acid sequence at least 95% identical to an aminoacid sequence selected from the group consisting of the LC CDR2sequences set forth in Table 5A; and (C) a light chain CDR3 comprising(i) an amino acid sequence selected from the group consisting of the LCCDR3 sequences set forth in Table 5A, (ii) an amino acid sequence atleast 90% identical to an amino acid sequence selected from the groupconsisting of the LC CDR3 sequences set forth in Table 5A, or (iii) anamino acid sequence at least 95% identical to an amino acid sequenceselected from the group consisting of the LC CDR3 sequences set forth inTable 5A.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising: (A) a heavy chain CDR1 comprising (i) an amino acid sequenceselected from the group consisting of the HC CDR1 sequences set forth inTable 5B, (ii) an amino acid sequence at least 90% identical to an aminoacid sequence selected from the group consisting of the HC CDR1sequences set forth in Table 5B, or (iii) an amino acid sequence atleast 95% identical to an amino acid sequence selected from the groupconsisting of the HC CDR1 sequences set forth in Table 5B; (B) a heavychain CDR2 comprising (i) an amino acid sequence selected from the groupconsisting of the HC CDR2 sequences set forth in Table 5B, (ii) an aminoacid sequence at least 90% identical to an amino acid sequence selectedfrom the group consisting of the HC CDR2 sequences set forth in Table5B, or (iii) an amino acid sequence at least 95% identical to an aminoacid sequence selected from the group consisting of the HC CDR2sequences set forth in Table 5B; and (C) a heavy chain CDR3 comprising(i) an amino acid sequence selected from the group consisting of the HCCDR3 sequences set forth in Table 5B, (ii) an amino acid sequence atleast 90% identical to an amino acid sequence selected from the groupconsisting of the HC CDR3 sequences set forth in Table 5B, or (iii) anamino acid sequence at least 95% identical to an amino acid sequenceselected from the group consisting of the HC CDR3 sequences set forth inTable 5B.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising (i) heavy chain CDRs CDR1, CDR2 and CDR3, each having asequence of the corresponding heavy chain CDR as above, and (ii) lightchain CDRs CDR1, CDR2 and CDR3, each having a sequence of thecorresponding heavy chain CDR as above.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising a light chain variable region comprising (i) an amino acidsequence selected from the group consisting of the V_(L) AA sequencesset forth in Table 4A, (ii) an amino acid sequence at least 90%identical to an amino acid sequence selected from the group consistingof the V_(L) AA sequences set forth in Table 4A, or (iii) an amino acidsequence at least 95% identical to an amino acid sequence selected fromthe group consisting of the V_(L) AA sequences set forth in Table 4A.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-8,CDRH2-2, and CDRH3-3; and light chain CDRs: CDRL1-3, CDRL2-2, andCDRL3-2. (A)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-8,CDRH2-2, and CDRH3-3; and light chain CDRs: CDRL1-3, CDRL2-2, andCDRL3-2. (B)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-11,CDRH2-5, and CDRH3-6; and light chain CDRs: CDRL1-1, CDRL2-1, andCDRL3-1. (C)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-1,CDRH2-4, and CDRH3-6; and light chain CDRs: CDRL1-2, CDRL2-1, andCDRL3-1. (D)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-1,CDRH2-4, and CDRH3-6; and light chain CDRs: CDRL1-1, CDRL2-1, andCDRL3-1. (E)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-1,CDRH2-4, and CDRH3-6; and light chain CDRs: CDRL1-1, CDRL2-1, andCDRL3-1. (F)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-1,CDRH2-4, and CDRH3-6; and light chain CDRs: CDRL1-1, CDRL2-1, andCDRL3-1. (G)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-1,CDRH2-4, and CDRH3-6; and light chain CDRs: CDRL1-2, CDRL2-1, andCDRL3-1. (H)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-3,CDRH2-5, and CDRH3-5; and light chain CDRs: CDRL1-1, CDRL2-1, andCDRL3-1. (J)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-3,CDRH2-5, and CDRH3-5; and light chain CDRs: CDRL1-1, CDRL2-1, andCDRL3-1. (L)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-9,CDRH2-6, and CDRH3-1 and light chain CDRs: CDRL1-4, CDRL2-3, andCDRL3-3. (M)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-9,CDRH2-6, and CDRH3-1; and light chain CDRs: CDRL1-4, CDRL2-3, andCDRL3-3. (N)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-4,CDRH2-7, and CDRH3-8; and light chain CDRs: CDRL1-6, CDRL2-4, andCDRL3-5. (O)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-4,CDRH2-7, and CDRH3-8; and light chain CDRs: CDRL1-6, CDRL2-4, andCDRL3-5. (P)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-4,CDRH2-7, and CDRH3-8; and light chain CDRs: CDRL1-6, CDRL2-4, andCDRL3-5. (Q)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-10,CDRH2-1, and CDRH3-7 and light chain CDRs: CDRL1-5, CDRL2-3, andCDRL3-4. (R)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-6,CDRH2-8, and CDRH3-2; and light chain CDRs: CDRL1-7, CDRL2-5, andCDRL3-6. (S)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-7,CDRH2-3, and CDRH3-4; and light chain CDRs: CDRL1-8, CDRL2-6, andCDRL3-7. (T)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-2,CDRH2-8, and CDRH3-2; and light chain CDRs: CDRL1-9, CDRL2-5, andCDRL3-8. (U)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-6,CDRH2-8, and CDRH3-2; and light chain CDRs: CDRL1-10, CDRL2-5, andCDRL3-6. (V)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-6,CDRH2-8, and CDRH3-2 and light chain CDRs: CDRL1-10, CDRL2-5, andCDRL3-6. (W)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-6,CDRH2-8, and CDRH3-2; and light chain CDRs: CDRL1-12, CDRL2-5, andCDRL3-6. (X)

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof, comprising heavy chain CDRs: CDRH1-5,CDRH2-8, and CDRH3-2; and light chain CDRs: CDRL1-11, CDRL2-5, andCDRL3-8. (Y)

In another aspect, the invention includes any of the foregoing (A, B, C,D, E, F, G, H, J, L, M, N, O, P, Q, R, S, T, U, V, W, X, or Y), whereinthe isolated antibody or antigen-binding fragment thereof specificallybinds human PAC1.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising a heavy chain CDR1 selected from the group consisting ofCDRH1-1, CDRH1-2, CDRH1-3, CDRH1-4, CDRH1-5, CDRH1-6, CDRH1-7, CDRH1-8,CDRH1-9, CDRH1-10, and CDRH1-11.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising a heavy chain CDR2 selected from the group consisting ofCDRH2-1, CDRH2-2, CDRH2-3, CDRH2-4, CDRH2-5, CDRH2-6, CDRH2-7, andCDRH2-8.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising a heavy chain CDR3 selected from the group consisting ofCDRH3-1, CDRH3-2, CDRH3-3, CDRH3-4, CDRH3-5, CDRH3-6, CDRH3-7, andCDRH3-8.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising a light chain CDR1 selected from the group consisting ofCDRL1-1, CDRL1-2, CDRL1-3, CDRL1-4, CDRL1-5, CDRL1-6, CDRL1-7, CDRL1-8,CDRL1-9, CDRL1-10, CDRL1-11 and CDRL1-12.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising a light chain CDR2 selected from the group consisting ofCDRL2-1, CDRL2-2, CDRL2-3, CDRL2-4, CDRL2-5, and CDRL2-6.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising a light chain CDR3 selected from the group consisting ofCDRL3-1, CDRL3-2, CDRL3-3, CDRL3-4, CDRL3-5, CDRL3-6, CDRL3-7, andCDRL3-8.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising a heavy chain CDR1 as above, a heavy chain CDR2 as above, aheavy chain CDR3 as above, a light chain CDR1 as above, a light chainCDR2 as above, and a light chain CDR3 as above.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising a heavy chain variable region comprising (i) an amino acidsequence selected from the group consisting of the V_(H) AA sequencesset forth in Table 4B, (ii) an amino acid sequence at least 90%identical to an amino acid sequence selected from the group consistingof the V_(H) AA sequences set forth in Table 4B, or (iii) an amino acidsequence at least 95% identical to an amino acid sequence selected fromthe group consisting of the V_(H) AA sequences set forth in Table 4B.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising a light chain variable region amino acid sequence selectedfrom the group consisting of the light chain variable region amino acidsequences of any of antibodies A, B, C, D, E, F, G, H, J, L, M, N, O, P,Q, R, S, T, U, V, W, X, and Y.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising a (i) a sequence selected from the group consisting of thesequences set forth in Table 4A, or (ii) a sequence at least 90%identical to one of the sequences in set forth in Table 4A, or (iii) asequence at least 95% identical to one of the sequences in set forth inTable 4A.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising a heavy chain variable region amino acid sequence selectedfrom the group consisting of the heavy chain variable region amino acidsequences of any of antibodies A, B, C, D, E, F, G, H, J, L, M, N, O, P,Q, R, S, T, U, V, W, X, and Y.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising a (i) a sequence selected from the group consisting of thesequences set forth in Table 4B, or (ii) a sequence at least 90%identical to one of the sequences in set forth in Table 4B, or (iii) asequence at least 95% identical to one of the sequences in set forth inTable 4B.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising a light chain amino acid sequence selected from the groupconsisting of the light chain amino acid sequences of any of antibodiesA, B, C, D, E, F, G, H, J, L, M, N, O, P, Q, R, S, T, U, V, W, X, and Y.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising a (i) a sequence selected from the group consisting of thesequences set forth in Table 2A, or (ii) a sequence at least 90%identical to one of the sequences in set forth in Table 2A, or (iii) asequence at least 95% identical to one of the sequences in set forth inTable 2A.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising a heavy chain amino acid sequence selected from the groupconsisting of the heavy chain amino acid sequences of any of antibodiesA, B, C, D, E, F, G, H, J, L, M, N, O, P, Q, R, S, T, U, V, W, X, and Y.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising a (i) a sequence selected from the group consisting of thesequences set forth in Table 2A, or (ii) a sequence at least 90%identical to one of the sequences in set forth in Table 2A, or (iii) asequence at least 95% identical to one of the sequences in set forth inTable 2A.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising a (i) a sequence selected from the group consisting of thesequences set forth in Table 2B, or (ii) a sequence at least 90%identical to one of the sequences in set forth in Table 2B, or (iii) asequence at least 95% identical to one of the sequences in set forth inTable 2B.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising a (i) a sequence selected from the group consisting of thesequences set forth in Table 4A, or (ii) a sequence at least 90%identical to one of the sequences in set forth in Table 4A, or (iii) asequence at least 95% identical to one of the sequences in set forth inTable 4A.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof that specifically binds human PAC1,comprising a (i) a sequence selected from the group consisting of thesequences set forth in Table 4B, or (ii) a sequence at least 90%identical to one of the sequences in set forth in Table 4B, or (iii) asequence at least 95% identical to one of the sequences in set forth inTable 4B.

In another aspect, the invention includes an isolated antibody orantigen-binding fragment thereof of any of the foregoing, wherein theantibody is a polyclonal antibody.

In another aspect, the invention includes the isolated antibody of anyof the above, wherein the isolated antibody is selected from the groupconsisting of a monoclonal antibody, a Fab fragment, an Fab′ fragment,an F(ab′)2 fragment, an Fv fragment, a diabody, and a single chainantibody.

In another aspect, the invention includes the isolated antibody asabove, wherein the isolated antibody is a monoclonal antibody selectedfrom the group consisting of a fully human antibody, a humanizedantibody and a chimeric antibody.

In another aspect, the invention includes the isolated antibody asabove, wherein the monoclonal antibody is an IgG1-, IgG2-, IgG3-, orIgG4-type antibody.

In another aspect, the invention includes the isolated antibody asabove, wherein the monoclonal antibody is an aglycosylated IgG1antibody.

In another aspect, the invention includes the isolated antibody asabove, wherein the monoclonal antibody has an N297G mutation in itsheavy chain.

In another aspect, the invention includes the isolated antibody asabove, wherein the monoclonal antibody is selected from the groupconsisting of antibodies A, D, E, J and V.

In another aspect, the invention includes the isolated antibody of anyas above, wherein the antibody selectively inhibits hPAC1 relative toVPA1 and VPAC2.

In another aspect, the invention includes the isolated antibody asabove, wherein the selectivity ratio is greater than 100:1.

In another aspect, the invention includes the isolated antibody asabove, wherein the selectivity ratio is greater than 1000:1.

In another aspect, the invention includes an isolated polynucleotidethat encodes an antibody of any as above.

In another aspect, the invention includes an isolated nucleic acidencoding an antibody or antigen-binding fragment thereof thatspecifically binds human PAC1, comprising (i) a sequence selected fromthe group consisting of the sequences set forth in Table 1A, or (ii) asequence at least 90% identical to one of the sequences in set forth inTable 1A, or (iii) a sequence at least 95% identical to one of thesequences in set forth in Table 1A.

In another aspect, the invention includes an isolated nucleic acidencoding an antibody or antigen-binding fragment thereof thatspecifically binds human PAC1, comprising (i) a sequence selected fromthe group consisting of the sequences set forth in Table 1B, or (ii) asequence at least 90% identical to one of the sequences in set forth inTable 1B, or (iii) a sequence at least 95% identical to one of thesequences in set forth in Table 1B.

In another aspect, the invention includes an isolated nucleic acidencoding an antibody or antigen-binding fragment thereof thatspecifically binds human PAC1, comprising (i) a sequence selected fromthe group consisting of the sequences set forth in Table 3A, or (ii) asequence at least 90% identical to one of the sequences in set forth inTable 3A, or (iii) a sequence at least 95% identical to one of thesequences in set forth in Table 3A.

In another aspect, the invention includes an isolated nucleic acidencoding an antibody or antigen-binding fragment thereof thatspecifically binds human PAC1, comprising (i) a sequence selected fromthe group consisting of the sequences set forth in Table 3B, or (ii) asequence at least 90% identical to one of the sequences in set forth inTable 3B, or (iii) a sequence at least 95% identical to one of thesequences in set forth in Table 3B.

In another aspect, the invention includes an isolated polynucleotidehaving a nucleic acid sequence selected from the group consisting of thesequences set forth in Table 1A, (ii) a nucleic acid sequence at least80% identical to a nucleic acid sequence selected from the groupconsisting of the sequences set forth in Table 1A, (iii) a nucleic acidsequence at least 90% identical to a nucleic acid sequence selected fromthe group consisting of the sequences set forth in Table 1A, or (iv) anucleic acid sequence at least 95% identical to a nucleic acid sequenceselected from the group consisting of the sequences set forth in Table1A.

In another aspect, the invention includes an isolated polynucleotidehaving a nucleic acid sequence selected from the group consisting of thesequences set forth in Table 1B, (ii) a nucleic acid sequence at least80% identical to a nucleic acid sequence selected from the groupconsisting of the sequences set forth in Table 1B, (iii) a nucleic acidsequence at least 90% identical to a nucleic acid sequence selected fromthe group consisting of the sequences set forth in Table 1B, or (iv) anucleic acid sequence at least 95% identical to a nucleic acid sequenceselected from the group consisting of the sequences set forth in Table1B.

In another aspect, the invention includes an isolated polynucleotidehaving a nucleic acid sequence selected from the group consisting of thesequences set forth in Table 3A, (ii) a nucleic acid sequence at least80% identical to a nucleic acid sequence selected from the groupconsisting of the sequences set forth in Table 3A, (iii) a nucleic acidsequence at least 90% identical to a nucleic acid sequence selected fromthe group consisting of the sequences set forth in Table 3A, or (iv) anucleic acid sequence at least 95% identical to a nucleic acid sequenceselected from the group consisting of the sequences set forth in Table3A.

In another aspect, the invention includes an isolated polynucleotidehaving a nucleic acid sequence selected from the group consisting of thesequences set forth in Table 3B, (ii) a nucleic acid sequence at least80% identical to a nucleic acid sequence selected from the groupconsisting of the sequences set forth in Table 3B, (iii) a nucleic acidsequence at least 90% identical to a nucleic acid sequence selected fromthe group consisting of the sequences set forth in Table 3B, or (iv) anucleic acid sequence at least 95% identical to a nucleic acid sequenceselected from the group consisting of the sequences set forth in Table3B.

In another aspect, the invention includes an expression vectorcomprising an isolated nucleic acid or polynucleotide of any as above.

In another aspect, the invention includes a cell line transformed withexpression vector as above.

In another aspect, the invention includes a method of making an antibodyor antigen binding fragment thereof of any as above, comprisingpreparing the antibody or antigen binding fragment thereof from a hostcell as above that secretes the antibody or antigen binding fragmentthereof.

In another aspect, the invention includes a pharmaceutical compositioncomprising an antibody or antigen binding fragment thereof of any asabove and a pharmaceutically acceptable excipient.

In another aspect, the invention includes a method for treating acondition associated with PAC1 in a patient, comprising administering toa patient an effective amount of an antibody or antigen binding fragmentthereof of any as above.

In another aspect, the invention includes the method as above, whereinthe condition is headache.

In another aspect, the invention includes the method as above, whereinthe condition is migraine.

In another aspect, the invention includes the method as above, whereinthe migraine is episodic migraine.

In another aspect, the invention includes the method as above, whereinthe migraine is chronic migraine.

In another aspect, the invention includes the method of any as above,wherein the method comprises prophylactic treatment.

DETAILED DESCRIPTION

The section headings used herein are for organizational purposes onlyand are not to be construed as limiting the subject matter described.

Unless otherwise defined herein, scientific and technical terms used inconnection with the present application shall have the meanings that arecommonly understood by those of ordinary skill in the art. Further,unless otherwise required by context, singular terms shall includepluralities and plural terms shall include the singular.

Generally, nomenclatures used in connection with, and techniques of,cell and tissue culture, molecular biology, immunology, microbiology,genetics and protein and nucleic acid chemistry and hybridizationdescribed herein are those well known and commonly used in the art. Themethods and techniques of the present application are generallyperformed according to conventional methods well known in the art and asdescribed in various general and more specific references that are citedand discussed throughout the present specification unless otherwiseindicated. See, e.g., Sambrook et al., Molecular Cloning: A LaboratoryManual, 3rd ed., Cold Spring Harbor Laboratory Press, Cold SpringHarbor, N.Y. (2001), Ausubel et al., Current Protocols in MolecularBiology, Greene Publishing Associates (1992), and Harlow and LaneAntibodies: A Laboratory Manual Cold Spring Harbor Laboratory Press,Cold Spring Harbor, N.Y. (1990), which are incorporated herein byreference. Enzymatic reactions and purification techniques are performedaccording to manufacturer's specifications, as commonly accomplished inthe art or as described herein. The terminology used in connection with,and the laboratory procedures and techniques of, analytical chemistry,synthetic organic chemistry, and medicinal and pharmaceutical chemistrydescribed herein are those well known and commonly used in the art.Standard techniques can be used for chemical syntheses, chemicalanalyses, pharmaceutical preparation, formulation, and delivery, andtreatment of patients.

It should be understood that this invention is not limited to theparticular methodology, protocols, and reagents, etc., described hereinand as such may vary. The terminology used herein is for the purpose ofdescribing particular embodiments only, and is not intended to limit thescope of the present invention, which is defined solely by the claims.

Other than in the operating examples, or where otherwise indicated, allnumbers expressing quantities of ingredients or reaction conditions usedherein should be understood as modified in all instances by the term“about.” The term “about” when used in connection with percentagesmeans±1%.

Definitions

The term “polynucleotide” or “nucleic acid” includes bothsingle-stranded and double-stranded nucleotide polymers. The nucleotidescomprising the polynucleotide can be ribonucleotides ordeoxyribonucleotides or a modified form of either type of nucleotide.

An “isolated nucleic acid molecule” means a DNA or RNA of genomic, mRNA,cDNA, or synthetic origin or some combination thereof which is notassociated with all or a portion of a polynucleotide in which theisolated polynucleotide is found in nature, or is linked to apolynucleotide to which it is not linked in nature. For purposes of thisdisclosure, it should be understood that “a nucleic acid moleculecomprising” a particular nucleotide sequence does not encompass intactchromosomes. Isolated nucleic acid molecules “comprising” specifiednucleic acid sequences may include, in addition to the specifiedsequences, coding sequences for up to ten or even up to twenty otherproteins or portions thereof, or may include operably linked regulatorysequences that control expression of the coding region of the recitednucleic acid sequences, and/or may include vector sequences.

Unless specified otherwise, the left-hand end of any single-strandedpolynucleotide sequence discussed herein is the 5′ end; the left-handdirection of double-stranded polynucleotide sequences is referred to asthe 5′ direction. The direction of 5′ to 3′ addition of nascent RNAtranscripts is referred to as the transcription direction; sequenceregions on the DNA strand having the same sequence as the RNA transcriptthat are 5′ to the 5′ end of the RNA transcript are referred to as“upstream sequences;” sequence regions on the DNA strand having the samesequence as the RNA transcript that are 3′ to the 3′ end of the RNAtranscript are referred to as “downstream sequences.”

The term “control sequence” refers to a polynucleotide sequence that canaffect the expression and processing of coding sequences to which it isligated. The nature of such control sequences may depend upon the hostorganism. In particular embodiments, control sequences for prokaryotesmay include a promoter, a ribosomal binding site, and a transcriptiontermination sequence. For example, control sequences for eukaryotes mayinclude promoters comprising one or a plurality of recognition sites fortranscription factors, transcription enhancer sequences, andtranscription termination sequence. “Control sequences” can includeleader sequences and/or fusion partner sequences.

The term “vector” means any molecule or entity (e.g., nucleic acid,plasmid, bacteriophage or virus) used to transfer protein codinginformation into a host cell.

The term “expression vector” or “expression construct” refers to avector that is suitable for transformation of a host cell and containsnucleic acid sequences that direct and/or control (in conjunction withthe host cell) expression of one or more heterologous coding regionsoperatively linked thereto. An expression construct may include, but isnot limited to, sequences that affect or control transcription,translation, and, if introns are present, affect RNA splicing of acoding region operably linked thereto.

As used herein, “operably linked” means that the components to which theterm is applied are in a relationship that allows them to carry outtheir inherent functions under suitable conditions. For example, acontrol sequence in a vector that is “operably linked” to a proteincoding sequence is ligated thereto so that expression of the proteincoding sequence is achieved under conditions compatible with thetranscriptional activity of the control sequences.

The term “host cell” means a cell that has been transformed, or iscapable of being transformed, with a nucleic acid sequence and therebyexpresses a gene of interest. The term includes the progeny of theparent cell, whether or not the progeny is identical in morphology or ingenetic make-up to the original parent cell, so long as the gene ofinterest is present.

The term “transduction” means the transfer of genes from one bacteriumto another, usually by bacteriophage. “Transduction” also refers to theacquisition and transfer of eukaryotic cellular sequences by replicationdefective retroviruses.

The term “transfection” means the uptake of foreign or exogenous DNA bya cell, and a cell has been “transfected” when the exogenous DNA hasbeen introduced inside the cell membrane. A number of transfectiontechniques are well known in the art and are disclosed herein. See,e.g., Graham et al., 1973, Virology 52:456; Sambrook et al., 2001,Molecular Cloning: A Laboratory Manual, supra; Davis et al., 1986, BasicMethods in Molecular Biology, Elsevier; Chu et al., 1981, Gene 13:197.Such techniques can be used to introduce one or more exogenous DNAmoieties into suitable host cells.

The term “transformation” refers to a change in a cell's geneticcharacteristics, and a cell has been transformed when it has beenmodified to contain new DNA or RNA. For example, a cell is transformedwhere it is genetically modified from its native state by introducingnew genetic material via transfection, transduction, or othertechniques. Following transfection or transduction, the transforming DNAmay recombine with that of the cell by physically integrating into achromosome of the cell, or may be maintained transiently as an episomalelement without being replicated, or may replicate independently as aplasmid. A cell is considered to have been “stably transformed” when thetransforming DNA is replicated with the division of the cell.

The terms “polypeptide” or “protein” are used interchangeably herein torefer to a polymer of amino acid residues. The terms also apply to aminoacid polymers in which one or more amino acid residues is an analog ormimetic of a corresponding naturally occurring amino acid, as well as tonaturally occurring amino acid polymers. The terms can also encompassamino acid polymers that have been modified, e.g., by the addition ofcarbohydrate residues to form glycoproteins, or phosphorylated.Polypeptides and proteins can be produced by a naturally-occurring andnon-recombinant cell; or it is produced by a genetically-engineered orrecombinant cell, and comprise molecules having the amino acid sequenceof the native protein, or molecules having deletions from, additions to,and/or substitutions of one or more amino acids of the native sequence.The terms “polypeptide” and “protein” specifically encompass antibodies,e.g., anti-PAC1 antibodies (aka PAC1 antibodies), PAC1 binding proteins,antibodies, or sequences that have deletions from, additions to, and/orsubstitutions of one or more amino acids of an antigen-binding protein.The term “polypeptide fragment” refers to a polypeptide that has anamino-terminal deletion, a carboxyl-terminal deletion, and/or aninternal deletion as compared with the full-length protein. Suchfragments may also contain modified amino acids as compared with thefull-length protein. In certain embodiments, fragments are about five to500 amino acids long. For example, fragments may be at least 5, 6, 8,10, 14, 20, 50, 70, 100, 110, 150, 200, 250, 300, 350, 400, or 450 aminoacids long. Useful polypeptide fragments include immunologicallyfunctional fragments of antibodies, including binding domains. In thecase of a PAC1-binding antibody, useful fragments include but are notlimited to a CDR region, a variable domain of a heavy or light chain, aportion of an antibody chain or just its variable domain including twoCDRs, and the like.

The term “isolated protein” (e.g., isolated antibody), “isolatedpolypeptide” or “isolated antibody” means that a subject protein,polypeptide or antibody is free of most other proteins with which itwould normally be found and has been separated from at least about 50percent of polynucleotides, lipids, carbohydrates, or other materialswith which it is associated in nature. Typically, an “isolated protein”,“isolated polypeptide” or “isolated antibody” constitutes at least about5%, at least about 10%, at least about 25%, or at least about 50% of agiven sample. Genomic DNA, cDNA, mRNA or other RNA, of synthetic origin,or any combination thereof may encode such an isolated protein.Preferably, the isolated protein polypeptide or antibody issubstantially free from other proteins or other polypeptides or othercontaminants that are found in its natural environment that wouldinterfere with its therapeutic, diagnostic, prophylactic, research orother use.

The terms “human PAC1”, “human PAC₁”, “hPAC1” and “hPAC₁”, “human PAC1receptor”, “human PAC₁ receptor”, “hPAC1 receptor” and “hPAC₁ receptor”are used interchangeably and refer to the human pituitary adenylatecyclase-activating polypeptide type I receptor. hPAC1 is a 468 aminoacid protein designated as P41586 (PACR_HUMAN) in theUniProtKB/Swiss-Prot database and is encoded by the ADCYAP1R1 gene.PACAP-27 and PACAP-38 are the principal endogenous agonists of PAC1.Unless otherwise specified or clear from the context in which the termis used, “PAC1” refers to human hPAC1.

A “variant” of a polypeptide (e.g., an antigen binding protein, or anantibody) comprises an amino acid sequence wherein one or more aminoacid residues are inserted into, deleted from and/or substituted intothe amino acid sequence relative to another polypeptide sequence.Variants include fusion proteins.

A “derivative” of a polypeptide is a polypeptide (e.g., an antigenbinding protein, or an antibody) that has been chemically modified insome manner distinct from insertion, deletion, or substitution variants,e.g., via conjugation to another chemical moiety.

The term “naturally occurring” as used throughout the specification inconnection with biological materials such as polypeptides, nucleicacids, host cells, and the like, refers to materials which are found innature.

An antibody or antigen binding fragment thereof is said to “specificallybind” its target when the dissociation constant (K_(D)) is ≦10⁻⁶ M. Theantibody specifically binds the target antigen with “high affinity” whenthe K_(D) is ≦1×10⁻⁸ M. In one embodiment, the antibodies orantigen-binding fragments thereof will bind to PAC1, or human PAC1 witha K_(D)≦5×10⁻⁷; in another embodiment the antibodies or antigen-bindingfragments thereof will bind with a K_(D)≦1×10⁻⁷; in another embodimentthe antibodies or antigen-binding fragments thereof will bind with aK_(D)≦5×10⁻⁸; in another embodiment the antibodies or antigen-bindingfragments thereof will bind with a K_(D)≦1×10⁻⁸; in another embodimentthe antibodies or antigen-binding fragments thereof will bind with aK_(D)≦5×10⁻⁹; in another embodiment the antibodies or antigen-bindingfragments thereof will bind with a K_(D)≦1×10⁻⁹; in another embodimentthe antibodies or antigen-binding fragments thereof will bind with aK_(D)≦5×10⁻¹⁰; in another embodiment the antibodies or antigen-bindingfragments thereof will bind with a K_(D)≦1×10¹⁰.

An antibody, antigen binding fragment thereof or antigen binding protein“selectively inhibits” a specific receptor relative to other receptorswhen the IC50 of the antibody, antigen binding fragment thereof orantigen binding protein in an inhibition assay of the specific receptoris at least 50-fold lower than the IC50 in an inhibition assay ofanother “reference” receptor, e.g., a hVPAC1 or hVPAC2 receptor. The“selectivity ratio” is the IC50 of the reference receptor divided byIC50 of the specific receptor.

“Antigen binding region” means a protein, or a portion of a protein,that specifically binds a specified antigen. For example, that portionof an antibody that contains the amino acid residues that interact withan antigen and confer on the antibody its specificity and affinity forthe antigen is referred to as “antigen binding region.” An antigenbinding region typically includes one or more “complementary bindingregions” (“CDRs”). Certain antigen binding regions also include one ormore “framework” regions. A “CDR” is an amino acid sequence thatcontributes to antigen binding specificity and affinity. “Framework”regions can aid in maintaining the proper conformation of the CDRs topromote binding between the antigen binding region and an antigen.

In certain aspects, recombinant antibodies or antigen-binding fragmentsthereof that bind PAC1 protein, or human PAC1, are provided. In thiscontext, a “recombinant protein” is a protein made using recombinanttechniques, i.e., through the expression of a recombinant nucleic acidas described herein. Methods and techniques for the production ofrecombinant proteins are well known in the art.

The term “antibody” refers to an intact immunoglobulin of any isotype,or an antigen binding fragment thereof that can compete with the intactantibody for specific binding to the target antigen, and includes, forinstance, chimeric, humanized, fully human, and bispecific antibodies orantigen-binding fragments thereof. An “antibody” as such is a species ofan antigen binding protein. An intact antibody generally will compriseat least two full-length heavy chains and two full-length light chains,but in some instances may include fewer chains such as antibodiesnaturally occurring in camelids which may comprise only heavy chains.Antibodies or antigen-binding fragments thereof may be derived solelyfrom a single source, or may be “chimeric,” that is, different portionsof the antibody may be derived from two different antibodies asdescribed further below. The antibodies or binding fragments thereof maybe produced in hybridomas, by recombinant DNA techniques, or byenzymatic or chemical cleavage of intact antibodies. Unless otherwiseindicated, the term “antibody” includes, in addition to antibodiescomprising two full-length heavy chains and two full-length lightchains, derivatives, variants, fragments, and mutations thereof,examples of which are described below.

The term “light chain” includes a full-length light chain and fragmentsthereof having sufficient variable region sequence to confer bindingspecificity. A full-length light chain includes a variable regiondomain, V_(L), and a constant region domain, C_(L). The variable regiondomain of the light chain is at the amino-terminus of the polypeptide.Light chains include kappa chains and lambda chains.

The term “heavy chain” includes a full-length heavy chain and fragmentsthereof having sufficient variable region sequence to confer bindingspecificity. A full-length heavy chain includes a variable regiondomain, V_(H), and three constant region domains, C_(H)1. C_(H)2, andC_(H)3. The V_(H) domain is at the amino-terminus of the polypeptide,and the C_(H) domains are at the carboxyl-terminus, with the C_(H)3being closest to the carboxy-terminus of the polypeptide. Heavy chainsmay be of any isotype, including IgG (including IgG1, IgG2, IgG3 andIgG4 subtypes), IgA (including IgA1 and IgA2 subtypes), IgM and IgE. Inone embodiment, the heavy chain is an aglycosylated IgG1, e.g., an IgG1HC with an N297G mutation.

The term “signal sequence”, “leader sequence” or “signal peptide” refersto a short (3-60 amino acids long) peptide chain that directs thetransport of a protein. Signal peptides may also be called targetingsignals, signal sequences, transit peptides, or localization signals.Some signal peptides are cleaved from the protein by signal peptidaseafter the proteins are transported, such that the biologically activeform of the protein (e.g., an antibody as described herein) is thecleaved, shorter form. Accordingly, terms such as “antibody comprising aheavy chain . . . ”, “antibody comprising a light chain . . . ”, etc.,where the antibody is characterized as having a heavy and/or light chainwith a particular identified sequence, are understood to includeantibodies having the specific identified sequences, antibodies havingthe specific identified sequences except that the signal sequences arereplaced by different signal sequences, as well as antibodies having theidentified sequences, minus any signal sequences.

The term “antigen binding fragment” (or simply “fragment”) of anantibody or immunoglobulin chain (heavy or light chain), as used herein,comprises a portion (regardless of how that portion is obtained orsynthesized) of an antibody that lacks at least some of the amino acidspresent in a full-length chain but which is capable of specificallybinding to an antigen. Such fragments are biologically active in thatthey bind specifically to the target antigen and can compete with otherantibodies or antigen-binding fragments thereof, for specific binding toa given epitope. In one aspect, such a fragment will retain at least oneCDR present in the full-length light or heavy chain, and in someembodiments will comprise a single heavy chain and/or light chain orportion thereof. These biologically active fragments may be produced byrecombinant DNA techniques, or may be produced, e.g., by enzymatic orchemical cleavage of intact antibodies. Immunologically functionalimmunoglobulin fragments include, but are not limited to, Fab, Fab′,F(ab′)₂, Fv, domain antibodies and single-chain antibodies, and may bederived from any mammalian source, including but not limited to human,mouse, rat, camelid or rabbit. It is contemplated further that afunctional portion of the antibodies disclosed herein, for example, oneor more CDRs, could be covalently bound to a second protein or to asmall molecule to create a therapeutic agent directed to a particulartarget in the body, possessing bifunctional therapeutic properties, orhaving a prolonged serum half-life.

An “Fab fragment” is comprised of one light chain and the C_(H)1 andvariable regions of one heavy chain. The heavy chain of a Fab moleculecannot form a disulfide bond with another heavy chain molecule.

An “Fc” region contains two heavy chain fragments comprising the C_(H)1and C_(H)2 domains of an antibody. The two heavy chain fragments areheld together by two or more disulfide bonds and by hydrophobicinteractions of the C_(H)3 domains.

An “Fab′ fragment” contains one light chain and a portion of one heavychain that contains the V_(H) domain and the C_(H)1 domain and also theregion between the C_(H)1 and C_(H)2 domains, such that an interchaindisulfide bond can be formed between the two heavy chains of two Fab′fragments to form an F(ab′)₂ molecule.

An “F(ab′)₂ fragment” contains two light chains and two heavy chainscontaining a portion of the constant region between the C_(H)1 andC_(H)2 domains, such that an interchain disulfide bond is formed betweenthe two heavy chains. A F(ab′)₂ fragment thus is composed of two Fab′fragments that are held together by a disulfide bond between the twoheavy chains.

The “Fv region” comprises the variable regions from both the heavy andlight chains, but lacks the constant regions.

“Single-chain antibodies” are Fv molecules in which the heavy and lightchain variable regions have been connected by a flexible linker to forma single polypeptide chain, which forms an antigen-binding region.Single chain antibodies are discussed in detail in International PatentApplication Publication No. WO 88/01649 and U.S. Pat. Nos. 4,946,778 and5,260,203, the disclosures of which are incorporated by reference.

A “domain antibody” is an immunologically functional immunoglobulinfragment containing only the variable region of a heavy chain or thevariable region of a light chain. In some instances, two or more V_(H)regions are covalently joined with a peptide linker to create a bivalentdomain antibody. The two V_(H) regions of a bivalent domain antibody maytarget the same or different antigens.

A “bivalent antigen binding protein” or “bivalent antibody” comprisestwo antigen binding sites. In some instances, the two binding sites havethe same antigen specificities. Bivalent antibodies may be bispecific,see, infra.

A “multispecific antigen binding protein” or “multispecific antibody” isone that targets more than one antigen or epitope.

A “bispecific,” “dual-specific” or “bifunctional” antigen bindingprotein or antibody is a hybrid antigen binding protein or antibody,respectively, having two different antigen binding sites. Bispecificantibodies are a species of multispecific antigen binding protein ormultispecific antibody and may be produced by a variety of methodsincluding, but not limited to, fusion of hybridomas or linking of Fab′fragments. See, e.g., Songsivilai and Lachmann, 1990, Clin. Exp.Immunol. 79:315-321; Kostelny et al., 1992, J. Immunol. 148:1547-1553.The two binding sites of a bispecific antigen binding protein orantibody will bind to two different epitopes, which may reside on thesame or different protein targets.

The term “neutralizing antigen binding protein” or “neutralizingantibody” refers to an antigen binding protein or antibody,respectively, that binds to a ligand, prevents binding of the ligand toits binding partner and interrupts the biological response thatotherwise would result from the ligand binding to its binding partner.In assessing the binding and specificity of an antigen binding protein,e.g., an antibody or immunologically functional antigen binding fragmentthereof, an antibody or fragment will substantially inhibit binding of aligand to its binding partner when an excess of antibody reduces thequantity of binding partner bound to the ligand by at least about 20%,30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 97%, 99% or more (asmeasured in an in vitro competitive binding assay). In the case of aPAC1 binding protein, such a neutralizing molecule will diminish theability of PAC1 to bind PACAP, e.g., PACAP-27 or PACAP-38.

The term “antigen” or “immunogen” refers to a molecule or a portion of amolecule capable of being bound by a selective binding agent, such as anantigen binding protein (including, e.g., an antibody or immunologicalfunctional antigen binding fragment thereof), and additionally capableof being used in an animal to produce antibodies capable of binding tothat antigen. An antigen may possess one or more epitopes that arecapable of interacting with different antibodies or fragments thereof.

The term “epitope” is the portion of a molecule that is bound by anantigen binding protein (for example, an antibody). The term includesany determinant capable of specifically binding to an antigen bindingprotein, such as an antibody or to a T-cell receptor. An epitope can becontiguous or non-contiguous (e.g., amino acid residues that are notcontiguous to one another in the polypeptide sequence but that within incontext of the molecule are bound by the antigen binding protein). Incertain embodiments, epitopes may be mimetic in that they comprise athree dimensional structure that is similar to an epitope used togenerate the antibody, yet comprise none or only some of the amino acidresidues found in that epitope used to generate the antibody. Mostoften, epitopes reside on proteins, but in some instances may reside onother kinds of molecules, such as nucleic acids. Epitope determinantsmay include chemically active surface groupings of molecules such asamino acids, sugar side chains, phosphoryl or sulfonyl groups, and mayhave specific three dimensional structural characteristics, and/orspecific charge characteristics. Generally, antibodies specific for aparticular target antigen will preferentially recognize an epitope onthe target antigen in a complex mixture of proteins and/ormacromolecules.

The term “identity” refers to a relationship between the sequences oftwo or more polypeptide molecules or two or more nucleic acid molecules,as determined by aligning and comparing the sequences. “Percentidentity” means the percent of identical residues between the aminoacids or nucleotides in the compared molecules and is calculated basedon the size of the smallest of the molecules being compared. For thesecalculations, gaps in alignments (if any) must be addressed by aparticular mathematical model or computer program (i.e., an“algorithm”). Methods that can be used to calculate the identity of thealigned nucleic acids or polypeptides include those described inComputational Molecular Biology, (Lesk, A. M., ed.), 1988, New York:Oxford University Press; Biocomputing Informatics and Genome Projects,(Smith, D. W., ed.), 1993, New York: Academic Press; Computer Analysisof Sequence Data, Part I, (Griffin, A. M., and Griffin, H. G., eds.),1994, New Jersey: Humana Press; von Heinje, G., 1987, Sequence Analysisin Molecular Biology, New York: Academic Press; Sequence AnalysisPrimer, (Gribskov, M. and Devereux, J., eds.), 1991, New York: M.Stockton Press; and Carillo et al., 1988, SIAM J. Applied Math. 48:1073.

In calculating percent identity, the sequences being compared arealigned in a way that gives the largest match between the sequences. Thecomputer program used to determine percent identity is the GCG programpackage, which includes GAP (Devereux et al., 1984, Nucl. Acid Res.12:387; Genetics Computer Group, University of Wisconsin, Madison,Wis.). The computer algorithm GAP is used to align the two polypeptidesor polynucleotides for which the percent sequence identity is to bedetermined. The sequences are aligned for optimal matching of theirrespective amino acid or nucleotide (the “matched span”, as determinedby the algorithm). A gap opening penalty (which is calculated as 3× theaverage diagonal, wherein the “average diagonal” is the average of thediagonal of the comparison matrix being used; the “diagonal” is thescore or number assigned to each perfect amino acid match by theparticular comparison matrix) and a gap extension penalty (which isusually 1/10 times the gap opening penalty), as well as a comparisonmatrix such as PAM 250 or BLOSUM 62 are used in conjunction with thealgorithm. In certain embodiments, a standard comparison matrix (see,Dayhoff et al., 1978, Atlas of Protein Sequence and Structure 5:345-352for the PAM 250 comparison matrix; Henikoff et al., 1992, Proc. Natl.Acad. Sci. U.S.A. 89:10915-10919 for the BLOSUM 62 comparison matrix) isalso used by the algorithm.

Recommended parameters for determining percent identity for polypeptidesor nucleotide sequences using the GAP program are the following:

Algorithm: Needleman et al., 1970, J. Mol. Biol. 48:443-453;

Comparison matrix: BLOSUM 62 from Henikoff et al., 1992, supra;

Gap Penalty: 12 (but with no penalty for end gaps)

Gap Length Penalty: 4

Threshold of Similarity: 0

Certain alignment schemes for aligning two amino acid sequences mayresult in matching of only a short region of the two sequences, and thissmall aligned region may have very high sequence identity even thoughthere is no significant relationship between the two full-lengthsequences. Accordingly, the selected alignment method (GAP program) canbe adjusted if so desired to result in an alignment that spans at least50 contiguous amino acids of the target polypeptide.

As used herein, “substantially pure” means that the described species ofmolecule is the predominant species present, that is, on a molar basisit is more abundant than any other individual species in the samemixture. In certain embodiments, a substantially pure molecule is acomposition wherein the object species comprises at least 50% (on amolar basis) of all macromolecular species present. In otherembodiments, a substantially pure composition will comprise at least80%, 85%, 90%, 95%, or 99% of all macromolecular species present in thecomposition. In other embodiments, the object species is purified toessential homogeneity wherein contaminating species cannot be detectedin the composition by conventional detection methods and thus thecomposition consists of a single detectable macromolecular species.

The term “treating” refers to any indicia of success in the treatment oramelioration of an injury, pathology or condition, including anyobjective or subjective parameter such as abatement; remission;diminishing of symptoms or making the injury, pathology or conditionmore tolerable to the patient; slowing in the rate of degeneration ordecline; making the final point of degeneration less debilitating;improving a patient's physical or mental well-being. The treatment oramelioration of symptoms can be based on objective or subjectiveparameters; including the results of a physical examination,neuropsychiatric exams, and/or a psychiatric evaluation. For example,certain methods presented herein successfully treat migraine headacheseither prophylactically or as an acute treatment, decreasing thefrequency of migraine headaches, decreasing the severity of migraineheadaches, and/or ameliorating a symptom associated with migraineheadaches.

An “effective amount” is generally an amount sufficient to reduce theseverity and/or frequency of symptoms, eliminate the symptoms and/orunderlying cause, prevent the occurrence of symptoms and/or theirunderlying cause, and/or improve or remediate the damage that resultsfrom or is associated with migraine headache. In some embodiments, theeffective amount is a therapeutically effective amount or aprophylactically effective amount. A “therapeutically effective amount”is an amount sufficient to remedy a disease state (e.g. migraineheadache) or symptoms, particularly a state or symptoms associated withthe disease state, or otherwise prevent, hinder, retard or reverse theprogression of the disease state or any other undesirable symptomassociated with the disease in any way whatsoever. A “prophylacticallyeffective amount” is an amount of a pharmaceutical composition that,when administered to a subject, will have the intended prophylacticeffect, e.g., preventing or delaying the onset (or reoccurrence) ofmigraine headache, or reducing the likelihood of the onset (orreoccurrence) of migraine headache or migraine headache symptoms. Thefull therapeutic or prophylactic effect does not necessarily occur byadministration of one dose, and may occur only after administration of aseries of doses. Thus, a therapeutically or prophylactically effectiveamount may be administered in one or more administrations.

“Amino acid” includes its normal meaning in the art. The twentynaturally-occurring amino acids and their abbreviations followconventional usage. See, Immunology-A Synthesis, 2nd Edition, (E. S.Golub and D. R. Green, eds.), Sinauer Associates: Sunderland, Mass.(1991), incorporated herein by reference for any purpose. Stereoisomers(e.g., D-amino acids) of the twenty conventional amino acids, unnaturalamino acids such as α-,α-disubstituted amino acids, N-alkyl amino acids,and other unconventional amino acids may also be suitable components forpolypeptides and are included in the phrase “amino acid.” Examples ofunconventional amino acids include: 4-hydroxyproline,γ-carboxyglutamate, ε-N,N,N-trimethyllysine, ε-N-acetyllysine,O-phosphoserine, N-acetylserine, N-formylmethionine, 3-methylhistidine,5-hydroxylysine, σ-N-methylarginine, and other similar amino acids andimino acids (e.g., 4-hydroxyproline). In the polypeptide notation usedherein, the left-hand direction is the amino terminal direction and theright-hand direction is the carboxyl-terminal direction, in accordancewith standard usage and convention.

PAC1 in Disease

Neuropeptides present in the perivascular space of cranial vessels havebeen implicated as important mediators of nociceptive input duringmigraine attacks. Pituitary adenylate cyclase-activating polypeptide(PACAP) is present in sensory trigeminal neurons and may modulatenociception at different levels of the nervous system. Humanexperimental studies have shown that PACAP38 induces both headache andmigraine-like attacks (Schytz, et al., 2009, “PACAP38 inducesmigraine-like attacks in patients with migraine without aura”, Brain:132pp 16-25), supporting the idea that PAC1 receptor antagonists may beused in the prophylactic and/or acute treatment of migraine.

Antibodies

Antibodies that bind PAC1 protein, including human PAC1 (hPAC1) proteinare provided herein. The antibodies provided are polypeptides into whichone or more complementary determining regions (CDRs), as describedherein, are embedded and/or joined. In some antibodies, the CDRs areembedded into a “framework” region, which orients the CDR(s) such thatthe proper antigen binding properties of the CDR(s) is achieved. Ingeneral, antibodies that are provided can interfere with, block, reduceor modulate the interaction between PAC1 and its ligand(s), e.g., PACAP,such as PACAP-38.

In certain embodiments, the antibodies include, but not limited to,monoclonal antibodies, bispecific antibodies, minibodies, domainantibodies, synthetic antibodies (sometimes referred to herein as“antibody mimetics”), chimeric antibodies, humanized antibodies, humanantibodies, antibody fusions (sometimes referred to herein as “antibodyconjugates”), and fragments thereof. The various structures are furtherdescribed herein below.

The antibodies provided herein have been demonstrated to bind to PAC1,in particular human PAC1 and cyno PAC1. As described further in theexamples below, certain antibodies were tested and found to bind toepitopes different from those bound by a number of other antibodiesdirected against one or the other of the components of PAC1. Theantibodies that are provided prevent PACAP (e.g., PACAP-38) from bindingto its receptor. As a consequence, the antibodies provided herein arecapable of inhibiting PAC1 activity. In particular, antibodies bindingto these epitopes can have one or more of the following activities:inhibiting, inter alia, induction of PAC1 signal transduction pathways,inhibiting vasodialation, causing vasoconstriction, decreasinginflammation, e.g., neurogenic inflammation, and other physiologicaleffects induced by PAC1 upon PACAP binding.

The antibodies that are disclosed herein have a variety of utilities.Some of the antibodies, for instance, are useful in specific bindingassays, affinity purification of PAC1, in particular hPAC1 or itsligands and in screening assays to identify other antagonists of PAC1activity. Some of the antibodies are useful for inhibiting binding of aPAC1 ligand (e.g., PACAP-38) to PAC1.

The antibodies can be used in a variety of treatment applications, asexplained herein. For example, certain PAC1 antibodies are useful fortreating conditions associated with PAC1 mediated signaling, such asreducing, alleviating, or treating the frequency and/or severity ofmigraine headache, reducing, alleviating, or treating cluster headache,reducing, alleviating, or treating chronic pain, alleviating or treatingdiabetes mellitus (type II), reducing, alleviating, or treatingcardiovascular disorders, and reducing, alleviating, or treatinghemodynamic derangements associated with endotoxemia and sepsis in apatient. Other uses for the antibodies include, for example, diagnosisof PAC1-associated diseases or conditions and screening assays todetermine the presence or absence of PAC1. Some of the antibodiesdescribed herein are useful in treating consequences, symptoms, and/orthe pathology associated with PAC1 activity. These include, but are notlimited to, various types of headaches, including migraine (e.g.,chronic and/or episodic migraine).

Some of the antibodies that are provided have the structure typicallyassociated with naturally occurring antibodies. The structural units ofthese antibodies typically comprise one or more tetramers, each composedof two identical couplets of polypeptide chains, though some species ofmammals also produce antibodies having only a single heavy chain. In atypical antibody, each pair or couplet includes one full-length “light”chain (in certain embodiments, about 25 kDa) and one full-length “heavy”chain (in certain embodiments, about 50-70 kDa). Each individualimmunoglobulin chain is composed of several “immunoglobulin domains”,each consisting of roughly 90 to 110 amino acids and expressing acharacteristic folding pattern. These domains are the basic units ofwhich antibody polypeptides are composed. The amino-terminal portion ofeach chain typically includes a variable domain that is responsible forantigen recognition. The carboxy-terminal portion is more conservedevolutionarily than the other end of the chain and is referred to as the“constant region” or “C region”. Human light chains generally areclassified as kappa and lambda light chains, and each of these containsone variable domain and one constant domain. Heavy chains are typicallyclassified as mu, delta, gamma, alpha, or epsilon chains, and thesedefine the antibody's isotype as IgM, IgD, IgG, IgA, and IgE,respectively. IgG has several subtypes, including, but not limited to,IgG1, IgG2, IgG3, and IgG4. IgM subtypes include IgM, and IgM2. IgAsubtypes include IgA1 and IgA2. In humans, the IgA and IgD isotypescontain four heavy chains and four light chains; the IgG and IgEisotypes contain two heavy chains and two light chains; and the IgMisotype contains five heavy chains and five light chains. The heavychain C region typically comprises one or more domains that may beresponsible for effector function. The number of heavy chain constantregion domains will depend on the isotype. IgG heavy chains, forexample, each contain three C region domains known as C_(H)1, C_(H)2 andC_(H)3. The antibodies that are provided can have any of these isotypesand subtypes. In certain embodiments, the PAC1 antibody is of the IgG1,IgG2, or IgG4 subtype.

In full-length light and heavy chains, the variable and constant regionsare joined by a “J” region of about twelve or more amino acids, with theheavy chain also including a “D” region of about ten more amino acids.See, e.g., Fundamental Immunology, 2nd ed., Ch. 7 (Paul, W., ed.) 1989,New York: Raven Press (hereby incorporated by reference in its entiretyfor all purposes). The variable regions of each light/heavy chain pairtypically form the antigen binding site.

Variable regions of immunoglobulin chains generally exhibit the sameoverall structure, comprising relatively conserved framework regions(FR) joined by three hypervariable regions, more often called“complementarity determining regions” or CDRs. The CDRs from the twochains of each heavy chain/light chain pair mentioned above typicallyare aligned by the framework regions to form a structure that bindsspecifically with a specific epitope on the target protein (e.g., PAC1).From N-terminal to C-terminal, naturally-occurring light and heavy chainvariable regions both typically conform with the following order ofthese elements: FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. A numberingsystem has been devised for assigning numbers to amino acids that occupypositions in each of these domains. This numbering system is defined inKabat Sequences of Proteins of Immunological Interest (1987 and 1991,NIH, Bethesda, Md.), or Chothia & Lesk, 1987, J. Mol. Biol. 196:901-917;Chothia et al., 1989, Nature 342:878-883.

The various heavy chain and light chain variable regions provided hereinare depicted in Tables 4A and 4B. Each of these variable regions may beattached to the above heavy and light chain constant regions to form acomplete antibody heavy and light chain, respectively. Further, each ofthe so generated heavy and light chain sequences may be combined to forma complete antibody structure. It should be understood that the heavychain and light chain variable regions provided herein can also beattached to other constant domains having different sequences than theexemplary sequences listed above.

Nucleic acids that encode for the antibodies described herein, orportions thereof, are also provided, including nucleic acids encodingone or both chains of an antibody, or a fragment, derivative, mutein, orvariant thereof, polynucleotides encoding heavy chain variable regionsor only CDRs, polynucleotides sufficient for use as hybridizationprobes, PCR primers or sequencing primers for identifying, analyzing,mutating or amplifying a polynucleotide encoding a polypeptide,anti-sense nucleic acids for inhibiting expression of a polynucleotide,and complementary sequences of the foregoing. The nucleic acids can beany length. They can be, for example, 5, 10, 15, 20, 25, 30, 35, 40, 45,50, 75, 100, 125, 150, 175, 200, 250, 300, 350, 400, 450, 500, 750,1,000, 1,500 or more nucleotides in length, and/or can comprise one ormore additional sequences, for example, regulatory sequences, and/or bepart of a larger nucleic acid, for example, a vector. The nucleic acidscan be single-stranded or double-stranded and can comprise RNA and/orDNA nucleotides, and artificial variants thereof (e.g., peptide nucleicacids).

Tables 1A, 1B, 3A and 3B show exemplary nucleic acid sequences. Anyvariable region provided herein may be attached to these constantregions to form complete heavy and light chain sequences. However, itshould be understood that these constant regions sequences are providedas specific examples only—one of skill in the art may employ otherconstant regions, including IgG1 heavy chain constant region, IgG3 orIgG4 heavy chain constant regions, any of the seven lambda light chainconstant regions, including hCL-1, hCL-2, hCL-3 and hCL-7; constantregions that have been modified for improved stability, expression,manufacturability or other desired characteristics, and the like. Insome embodiments, the variable region sequences are joined to otherconstant region sequences that are known in the art. Exemplary nucleicacid sequences encoding heavy and light chain variable regions areprovided in Tables 3A and 3B.

Specific examples of full length light and heavy chains of theantibodies that are provided and their corresponding nucleic and aminoacid sequences are summarized in Tables 1A, 1B, 2A and 2B. Tables 1A and2A show exemplary light chain sequences, and Tables 1B and 2B showexemplary heavy chain sequences, which are shown without the respectivesignal sequences.

TABLE 1A  Exemplary Anti-hPAC1 Antibody Light ChainNucleic Acid Sequences SEQ ID Ab NO: ID LC NA Sequence 1 AGACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAATCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGGTATTTAAATTGGTATCAACAGAAACCAGGGAAAGCCCCTAAACTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGATCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAACAGTCTGCAACCTGAAGATTTTGCAACTTACTTCTGTCAACAGAGTTACAGTCCCCCATTCACTTTCGGCCCTGGGACCAAAGTGGATATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT 1 BGACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAATCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGGTATTTAAATTGGTATCAACAGAAACCAGGGAAAGCCCCTAAACTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGATCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAACAGTCTGCAACCTGAAGATTTTGCAACTTACTTCTGTCAACAGAGTTACAGTCCCCCATTCACTTTCGGCCCTGGGACCAAAGTGGATATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT 3 CGATATCCAGCTCACTCAATCGCCATCATTTCTCTCCGCTTCGGTAGGCGACCGGGTCACGATCACATGCAGGGCGTCGCAAAGCATTGGGAGGTCGTTGCATTGGTATCAGCAGAAACCCGGAAAGGCCCCGAAACTTCTGATCAAATACGCATCACAAAGCTTGAGCGGTGTGCCGTCGCGCTTCTCCGGTTCCGGAAGCGGAACGGAATTCACGCTTACAATCTCCTCACTGCAGCCCGAGGATTTCGCGACCTATTACTGTCACCAGTCATCCAGACTCCCGTTTACTTTTGGCCCTGGGACCAAGGTGGACATTAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT 5 DGAGATCGTACTTACTCAGTCACCCGCCACATTGTCCCTGAGCCCGGGTGAACGGGCGACCCTCAGCTGCCGAGCATCCCAGTCCGTCGGACGATCATTGCACTGGTACCAACAAAAACCGGGCCAGGCCCCCAGACTTCTGATCAAGTATGCGTCACAGAGCTTGTCGGGTATTCCCGCTCGCTTTTCGGGGTCGGGATCCGGGACAGATTTCACGCTCACAATCTCCTCGCTGGAACCCGAGGACTTCGCGGTCTACTATTGTCATCAGTCATCGAGGTTGCCTTTCACGTTTGGACCAGGGACCAAGGTGGACATTAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT 7 EGATATCCAGCTCACTCAATCGCCATCATTTCTCTCCGCTTCGGTAGGCGACCGGGTCACGATCACATGCAGGGCGTCGCAAAGCATTGGGAGGTCGTTGCATTGGTATCAGCAGAAACCCGGAAAGGCCCCGAAACTTCTGATCAAATACGCATCACAAAGCTTGAGCGGTGTGCCGTCGCGCTTCTCCGGTTCCGGAAGCGGAACGGAATTCACGCTTACAATCTCCTCACTGCAGCCCGAGGATTTCGCGACCTATTACTGTCACCAGTCATCCAGACTCCCGTTTACTTTTGGCCCTGGGACCAAGGTGGACATTAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT 7 FGATATCCAGCTCACTCAATCGCCATCATTTCTCTCCGCTTCGGTAGGCGACCGGGTCACGATCACATGCAGGGCGTCGCAAAGCATTGGGAGGTCGTTGCATTGGTATCAGCAGAAACCCGGAAAGGCCCCGAAACTTCTGATCAAATACGCATCACAAAGCTTGAGCGGTGTGCCGTCGCGCTTCTCCGGTTCCGGAAGCGGAACGGAATTCACGCTTACAATCTCCTCACTGCAGCCCGAGGATTTCGCGACCTATTACTGTCACCAGTCATCCAGACTCCCGTTTACTTTTGGCCCTGGGACCAAGGTGGACATTAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT 7 GGATATCCAGCTCACTCAATCGCCATCATTTCTCTCCGCTTCGGTAGGCGACCGGGTCACGATCACATGCAGGGCGTCGCAAAGCATTGGGAGGTCGTTGCATTGGTATCAGCAGAAACCCGGAAAGGCCCCGAAACTTCTGATCAAATACGCATCACAAAGCTTGAGCGGTGTGCCGTCGCGCTTCTCCGGTTCCGGAAGCGGAACGGAATTCACGCTTACAATCTCCTCACTGCAGCCCGAGGATTTCGCGACCTATTACTGTCACCAGTCATCCAGACTCCCGTTTACTTTTGGCCCTGGGACCAAGGTGGACATTAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT 5 HGAGATCGTACTTACTCAGTCACCCGCCACATTGTCCCTGAGCCCGGGTGAACGGGCGACCCTCAGCTGCCGAGCATCCCAGTCCGTCGGACGATCATTGCACTGGTACCAACAAAAACCGGGCCAGGCCCCCAGACTTCTGATCAAGTATGCGTCACAGAGCTTGTCGGGTATTCCCGCTCGCTTTTCGGGGTCGGGATCCGGGACAGATTTCACGCTCACAATCTCCTCGCTGGAACCCGAGGACTTCGCGGTCTACTATTGTCATCAGTCATCGAGGTTGCCTTTCACGTTTGGACCAGGGACCAAGGTGGACATTAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT 9 JGATATCCAGCTCACTCAATCGCCATCATTTCTCTCCGCTTCGGTAGGCGACCGGGTCACGATCACATGCAGGGCGTCGCAAAGCATTGGGAGGTCGTTGCATTGGTATCAGCAGAAACCCGGAAAGGCCCCGAAACTTCTGTTCAAATACGCATCACAAAGCTTGAGCGGTGTGCCGTCGCGCTTCTCCGGTTCCGGAAGCGGAACGGAATTCACGCTTACAATCTCCTCACTGCAGCCCGAGGATTTCGCGACCTATTACTGTCACCAGTCATCCAGACTCCCGTTTACTTTTGGCCCTGGGACCAAGGTGGACATTAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT 9 LGATATCCAGCTCACTCAATCGCCATCATTTCTCTCCGCTTCGGTAGGCGACCGGGTCACGATCACATGCAGGGCGTCGCAAAGCATTGGGAGGTCGTTGCATTGGTATCAGCAGAAACCCGGAAAGGCCCCGAAACTTCTGTTCAAATACGCATCACAAAGCTTGAGCGGTGTGCCGTCGCGCTTCTCCGGTTCCGGAAGCGGAACGGAATTCACGCTTACAATCTCCTCACTGCAGCCCGAGGATTTCGCGACCTATTACTGTCACCAGTCATCCAGACTCCCGTTTACTTTTGGCCCTGGGACCAAGGTGGACATTAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT 11 MGAAATTGTGTTGACGCAGTCGCCAGGCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTAGCAGCAGCTACTTAGCCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGGTGCATCCAGCAGGGCCACTGGCATCCCAGACAGGTTCAGTAACAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGACTGGAGCCTGAAGATTTTGCAGTGTATTACTGTCAGAGGTATGGTAGCTCACGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGTACCGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT 11 NGAAATTGTGTTGACGCAGTCGCCAGGCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTAGCAGCAGCTACTTAGCCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGGTGCATCCAGCAGGGCCACTGGCATCCCAGACAGGTTCAGTAACAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGACTGGAGCCTGAAGATTTTGCAGTGTATTACTGTCAGAGGTATGGTAGCTCACGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGTACCGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT 13 OGATATTGTGATGACTCAGTCTCCACTCTCCCTGCCCGTCACCCCTGGAGAGCCGGCCTCCATCTCCTGCAGGTCTAGTCAGAGCCTCCTGCATAGTAATGGATACAACTATTTGGATTGGTACCTGCAGAAGCCAGGGCAGTCTCCACAGCTCCTGCTCTATTTGGGTTCTAATCGGGCCTCCGGGGTCCCTGACAGGTTCAGTGGCAGTGGATCAGGCACAGATTTTACACTGCAAATCAGCAGAGTGGAGGCTGAGGATGTTGGGGTTTATTACTGCATGCAAACTCTACAAACTCCATTCACTTTCGGCCCTGGGACCAAAGTGGATATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGG GGAGAGTGT 15 PGATATTGTGATGACTCAGTCTCCACTCTCCCTGCCCGTCACCCCTGGAGAGCCGGCCTCCATCTCCTGCAGGTCTAGTCAGAGCCTCCTGCATAGTAATGGATACAACTATTTGGATTGGTACCTGCAGAAGCCAGGGCAGTCTCCACAGCTCCTGCTCTATTTGGGTTCTAATCGGGCCTCCGGGGTCCCTGACAGGTTCAGTGGCAGTGGATCAGGCACAGATTTTACACTGAAAATCAGCAGAGTGGAGGCTGAGGATGTTGGGGTTTATTACTGCATGCAAACTCTACAAACTCCATTCACTTTCGGCCCTGGGACCAAAGTGGATATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGG GGAGAGTGT 15 QGATATTGTGATGACTCAGTCTCCACTCTCCCTGCCCGTCACCCCTGGAGAGCCGGCCTCCATCTCCTGCAGGTCTAGTCAGAGCCTCCTGCATAGTAATGGATACAACTATTTGGATTGGTACCTGCAGAAGCCAGGGCAGTCTCCACAGCTCCTGCTCTATTTGGGTTCTAATCGGGCCTCCGGGGTCCCTGACAGGTTCAGTGGCAGTGGATCAGGCACAGATTTTACACTGAAAATCAGCAGAGTGGAGGCTGAGGATGTTGGGGTTTATTACTGCATGCAAACTCTACAAACTCCATTCACTTTCGGCCCTGGGACCAAAGTGGATATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGG GGAGAGTGT 17 RGAAATTGTGTTGACGCAGTCTCCAGGCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGACTGTTAGCAGGAGCTACTTAGCCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGGTGCATCCAGCAGGGCCACTGGCATCCCAGACAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGACTGGAGCCTGAAGATTTTGCCGTGTTTTACTGTCAGCAGTTTGGTAGCTCACCGTGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT 19 SGACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCACCATCCATTGCAAGTCCAGCCAGAATGTTTTATACAGCTCCAACAATAAGAACTTCTTAACTTGGTACCAGCAGAAACCAGGACAGCCCCCTAAACTGCTCATTTACCGGGCATCTACCCGGGAATCCGGGGTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACGGATTTCACTCTCACTATCAGCAGTCTGCAGGCTGAAGATGTGGCAGTTTATTTCTGTCAGCAATATTATAGTGCTCCATTCACTTTCGGCCCTGGGACCAAAGTGGATATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAAC AGGGGAGAGTGT 21 TGACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGACCACCATCAAGTGCAAGTCCAGCCAGAGTGTTTTATACAGATCCAACAATAACAACTTCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTATTGGGCATCTACCCGGGAATCCGGGGTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGGCTGAAGATGTGGCTGTTTATTTCTGTCAGCAATATTATATTTCTCCGCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAAC AGGGGAGAGTGT 23 UGACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTATACAGTTCCAACAATAAGCACTACTTAGCTTGGTACCGGCAGAAACCAGGACAGCCTCCTAAACTGCTCATTTACAGGGCATCTACCCGGGAATCCGGGGTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTGGCAGTGTATTACTGTCAGCAATATTATAGTTCTCCATTCACTTTCGGCCCTGGGACCAAAGTGGATATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAA CAGGGGAGAGTGT 25 VGACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCACCATCCACTGCAAGTCCAGCCAGAGTGTTTTATACAGCTCCAACAATAAGAACTTCTTAACTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAACTTCTCATTTACCGGGCATCTACCCGGGAATCCGGGGTTCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGGCTGAAGATGTGGCAGTTTATTTCTGTCAGCAATATTATAGTGCTCCATTCACTTTCGGCCCTGGGACCAGAGTGGATATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAAC AGGGGAGAGTGT 25 WGACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCACCATCCACTGCAAGTCCAGCCAGAGTGTTTTATACAGCTCCAACAATAAGAACTTCTTAACTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAACTTCTCATTTACCGGGCATCTACCCGGGAATCCGGGGTTCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGGCTGAAGATGTGGCAGTTTATTTCTGTCAGCAATATTATAGTGCTCCATTCACTTTCGGCCCTGGGACCAGAGTGGATATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAAC AGGGGAGAGTGT 27 XGACATCGTGATGACTCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCACCATCCACTGCAAGTCCAGCCAGAGTGTTTTATACAGCTCCAACAATAGGAACTTCTTAAGTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAACTGCTCATTTACCGGGCATCTACCCGGGAATCCGGGGTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGGCTGAAGATGTGGCAGTTTATTTCTGTCAGCAATATTATAGTGCTCCATTCACTTTCGGCCCTGGGACCACAGTGGATATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAAC AGGGGAGAGTGT 29 YGACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTATACAGTTCCAACAATAAGAACTACTTAGCTTGGTACCGGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACAGGGCATCTACCCGGGAATCCGGGGTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGGCTGAAGATGTGGCAGTGTATCACTGTCAGCAATATTATAGTTCTCCATTCACTTTCGGCCCTGGGACCAAAGTGGATATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAA CAGGGGAGAGTGT

TABLE 1B  Exemplary Anti-hPAC1 Antibody Heavy ChainNucleic Acid Sequences SEQ ID Ab NO: ID HC NA Sequence 31 ACAGGTACAGCTGCAGCAGTCAGGTCCAGGACTGGTGAAGCCCTCGCAGACCCTCTCACTCACCTGTGCCATCTCCGGGGACAGTGTCTCTAGCAACAGTGCTACTTGGAACTGGATCAGGCAGTCCCCATCGAGAGGCCTTGAGTGGCTGGGAAGGACATATTACAGGTCCAAGTGGTCTAATCATTATGCAGTATCTGTGAAAAGTCGAATAACCATCAACCCCGACACGTCCAAGAGCCAGTTCTCCCTGCAGCTGAACTCTGTGACTCCCGAGGACACGGCTGTGTATTACTGTGCAAGAGGAACGTGGAAACAGCTATGGTTCCTTGACCACTGGGGCCAGGGAACCCTGGTCACCGTCTCTAGTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGGCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTC TCCGGGTAAA 33 BCAGGTACAGCTGCAGCAGTCAGGTCCAGGACTGGTGAAGCCCTCGCAGACCCTCTCACTCACCTGTGCCATCTCCGGGGACAGTGTCTCTAGCAACAGTGCTACTTGGAACTGGATCAGGCAGTCCCCATCGAGAGGCCTTGAGTGGCTGGGAAGGACATATTACAGGTCCAAGTGGTCTAATCATTATGCAGTATCTGTGAAAAGTCGAATAACCATCAACCCCGACACGTCCAAGAGCCAGTTCTCCCTGCAGCTGAACTCTGTGACTCCCGAGGACACGGCTGTGTATTACTGTGCAAGAGGAACGTGGAAACAGCTATGGTTCCTTGACCACTGGGGCCAGGGAACCCTGGTCACCGTCTCTAGTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA 35 CCAGGTGCAGTTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTTACTATGCCATACACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTAGAGTGGGTGGCAGTTATCTCATATGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTGTGTATTACTGTGCGAGAGGATACGATCTTTTGACTGGTTACCCCGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGC AGAAGAGCCTCTCCCTGTCTCCGGGTAAA37 D CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGCGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTAGATTTGCCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCATATGATGGAGGAAATAAATACTATGCAGAGTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTCTGTTTTACTGTGCGAGAGGATACGATGTTTTGACTGGTTACCCCGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCTAGTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGGCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTA AA 39 ECAAGTTCAGTTGGTGGAGTCTGGAGCCGAAGTAGTAAAGCCAGGAGCTTCAGTGAAAGTCTCTTGTAAAGCAAGTGGATTCACGTTTAGCCGCTTTGCCATGCATTGGGTGCGGCAAGCTCCCGGTCAGGGGTTGGAGTGGATGGGAGTTATTAGCTATGACGGGGGCAATAAGTACTACGCCGAGTCTGTTAAGGGTCGGGTCACAATGACACGGGACACCTCAACCAGTACACTCTATATGGAACTGTCTAGCCTGAGATCCGAGGACACCGCTGTGTATTATTGCGCTAGGGGGTACGATGTATTGACGGGTTATCCTGATTACTGGGGGCAGGGGACACTCGTAACCGTCTCTAGTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGGCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTA AA 41 FCAAGTTCAGTTGGTGGAGTCTGGAGCCGAAGTAGTAAAGCCAGGAGCTTCAGTGAAAGTCTCTTGTAAAGCAAGTGGATTCACGTTTAGCCGCTTTGCCATGCATTGGGTGCGGCAAGCTCCCGGTCAGGGGTTGGAGTGGATGGGAGTTATTAGCTATGACGGGGGCAATAAGTACTACGCCGAGTCTGTTAAGGGTCGGGTCACAATGACACGGGACACCTCAACCAGTACACTCTATATGGAACTGTCTAGCCTGAGATCCGAGGACACCGCTGTGTATTATTGCGCTAGGGGGTACGATGTATTGACGGGTTATCCTGATTACTGGGGGCAGGGGACACTCGTAACCGTCTCTAGTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGC AGAAGAGCCTCTCCCTGTCTCCGGGTAAA43 G CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGCGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTAGATTTGCCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCATATGATGGAGGAAATAAATACTATGCAGAGTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTCTGTTTTACTGTGCGAGAGGATACGATGTTTTGACTGGTTACCCCGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGC AGAAGAGCCTCTCCCTGTCTCCGGGTAAA43 H CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGCGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTAGATTTGCCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCATATGATGGAGGAAATAAATACTATGCAGAGTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTCTGTTTTACTGTGCGAGAGGATACGATGTTTTGACTGGTTACCCCGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGC AGAAGAGCCTCTCCCTGTCTCCGGGTAAA45 J CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGCGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTCGCTATGCCATGCACTGGGTCCGCCAGGCTTCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCATATGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTGCTAATGAGCAGCCTGAGAGCTGAGGACACGGCTGTGTTTTACTGTGCGAGAGGATACGATATTTTGACTGGTTACCCCGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCTAGTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGGCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTA AA 47 LCAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGCGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTCGCTATGCCATGCACTGGGTCCGCCAGGCTTCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCATATGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTGCTAATGAGCAGCCTGAGAGCTGAGGACACGGCTGTGTTTTACTGTGCGAGAGGATACGATATTTTGACTGGTTACCCCGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGC AGAAGAGCCTCTCCCTGTCTCCGGGTAAA49 M CAGGTTCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTCTCCTGCAAGGCTTCTGGTTACACCTTTACCAGCTATGGTATCAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGATGGATCAACGCTTACAATGGTCACACAAACTATGCACAGACGTTCCAGGGCAGAGTCACCATGACCACAGACACATCCACGAGCACAGCCTACATGGAGCTGAGGAGCCTGAGATCTGACGACACGGCCGTGTATTACTGTGCGAGGGAACTGGAACTACGCTCCTTCTATTACTTCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCCCCGTCTCTAGTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA 51 NCAGGTTCAGCTGGTGCAGTCTGGAGCTGAAGTGAAGAAGCCTGGGGCCTCAGTGAAGGTCTCCTGCAAGGCTTCTGGTTACACCTTTACCAGCTATGGTATCAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGATGGATCAACGCTTACAATGGTCACACAAACTATGCACAGACGTTCCAGGGCAGAGTCACCATGACCACAGACACATCCACGAGCACAGCCTACATGGAACTGAGGAGCCTGAGATCTGACGACACGGCCGTGTATTACTGTGCGAGGGAACTGGAACTACGCTCCTTCTATTACTTCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCTAGTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA 53 OCAGGTTCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGTCTGGGGCCTCTTTGAAGGTCTCCTGCAAGGCTTCTGGTTACATTTTTACCCGCTATGGTGTCAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGATGGATCACCACTTACAATGGTAACACAAACTATGCACAGAAGCTCCAGGGCAGAGTCACCATGACCATAGACACATCCACGAGCACAGCCTACATGGAACTGAGAAGCCTCAGATCTGACGACACGGCCGTGTATTACTGTGCGAGAAGAGTGCGGTATAGTGGGGGCTACTCGTTTGACAACTGGGGCCAGGGAACCCTGGTCACCGTCTCTAGTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA 55 PCAGGTTCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGTCTGGGGCCTCTTTGAAGGTCTCCTGCAAGGCTTCTGGTTACATTTTTACCCGCTATGGTGTCAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGATGGATCACCACTTACAATGGTAATACAAACTATGCACAGAAGCTCCAGGGCAGAGTCACCATGACCACAGACACATCCACGAGCACAGCCTACATGGAACTGAGGAGCCTCAGATCTGACGACACGGCCGTGTATTACTGTGCGAGAAGAGTGCGGTACAGTGGGGGCTACTCGTTTGACAACTGGGGCCAGGGAACCCTGGTCACCGTCTCTAGTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA 57 QCAGGTTCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGTCTGGGGCCTCTTTGAAGGTCTCCTGCAAGGCTTCTGGTTACATTTTTACCCGCTATGGTGTCAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGATGGATCACCACTTACAATGGTAATACAAACTATGCACAGAAACTCCAGGGCAGAGTCACCATGACCACAGACACATCCACGAACACAGCCTACATGGAACTGAGGAGCCTCAGATCTGACGACACGGCCGTGTATTACTGTGCGAGAAGAGTGCGGTATAGTGGGGGCTACTCGTTTGACAACTGGGGCCAGGGAACCCTGGTCACCGTCTCTAGTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA 59 RCAGGTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCGGAGACCCTGTCCCTCACCTGCACTGTCTCTGGTGGCTCCATCAGTAGTTACTACTGGAGCTGGATCCGGCAGCCCGCCGGGAAGGGACTGGAATGGATTGGGCGTATCTATACCAGTGGGAGCACCAACTACAACCCCTCCCTCAAGAGTCGAGTCACCATGTCAATAGGCACGTCCAAGAACCAGTTCTCCCTGAAGCTGAGCTCTGTGACCGCCGCGGACACGGCCGTGTATTACTGTGCGATTATTGCATCTCGTGGCTGGTACTTCGATCTCTGGGGCCGTGGCACCCTGGTCACCGTCTCTAGTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGC CTCTCCCTGTCTCCGGGTAAA 61 SCAGGTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCACAGACCCTGTCCCTCACCTGCACTGTCTCTGGTGGCTCCATCAGCAGTGGTGGTTACTACTGGAGCTGGATCCGCCAGCACCCAGGGAAGGGCCTGGAGTGGATTGGGTACATCTATTACAGTGGGAACACCTACTACAACCCGTCCCTCAAGAGTCGAGTTACCATATCAGGAGACACGTCTAAGAACCAGTTCTCCCTGAAGCTGAGGTCTGTGACTGCCGCGGACACGGCCGTGTATTACTGTGCGAGAGGAGGAGCAGCTCGCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCTAGTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGC AGAAGAGCCTCTCCCTGTCTCCGGGTAAA63 T CAGGTACAGCTGCAGCAGTCAGGTCCAGGACTGGTGAAGCCCTCGCAGACCCTCTCACTCACCTGTGCCATCTCCGGGGACAGTGTCTCTAGCAACAGTGCTGCTTGGAACTGGATCAGGCAGTCCCCATCGAGAGGCCTTGAGTGGCTGGGAAGGACATACTACAGGTCCAGGTGGTATAATGATTATGCAGTATCTGTGAAAAGTCGAATAACCATCAACCCAGACACATCCAAGAACCAGTTCTCCCTGCAGCTGAACTCTGTGACTCCCGAGGACACGGCTGTGTATTACTGTGCAAGAGGGGTCTTTTATAGCAAAGGTGCTTTTGATATCTGGGGCCAAGGGACAATGGTCACCGTCTCTAGTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA 65 UCAGGTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCACAGACCCTGTCCCTCACCTGCACTGTCTCTGGTGGCTCCATCAGCCGTGGTGGTTACTACTGGAGCTGGATCCGCCAGCACCCAGGGAAGGGCCTGGAGTGGATTGGGTACATATATTACAGTGGGAATACCTACTACAACCCGTCCCTCAAGAGTCGAGTTATCATATCAGGAGACACGTCTAAGAACCAGCTCTCCCTGAAGCTGAGGTCTGTGACTGCCGCGGACACGGCCGTGTATTATTGTGCGAGAGGAGGAGCAGCTCGCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCTAGTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAG AAGAGCCTCTCCCTGTCTCCGGGTAAA 67V CAGGTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCACAGACCCTGTCCCTCACCTGCACTGTCTCTGGTGGCTCCATCAGCAGTGGTGGTTACTACTGGAGCTGGATCCGCCAGCACCCAGGGAAGGGCCTGGAGTGGATTGGGTACATCTATTACAGTGGGAACACCTACTACAACCCGTCCCTCAAGAGTCGAGTTACCATATCAGGAGACACGTCTAAGAACCAGTTCTCCCTGAAGCTGAGGTCTGTGACTGCCGCGGACACGGCCGTGTATTACTGTACGAGAGGAGGAGCAGCTCGCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCTAGTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGGCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA 69 WCAGGTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCACAGACCCTGTCCCTCACCTGCACTGTCTCTGGTGGCTCCATCAGCAGTGGTGGTTACTACTGGAGCTGGATCCGCCAGCACCCAGGGAAGGGCCTGGAGTGGATTGGGTACATCTATTACAGTGGGAACACCTACTACAACCCGTCCCTCAAGAGTCGAGTTACCATATCAGGAGACACGTCTAAGAACCAGTTCTCCCTGAAGCTGAGGTCTGTGACTGCCGCGGACACGGCCGTGTATTACTGTACGAGAGGAGGAGCAGCTCGCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCTAGTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGC AGAAGAGCCTCTCCCTGTCTCCGGGTAAA61 X CAGGTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCACAGACCCTGTCCCTCACCTGCACTGTCTCTGGTGGCTCCATCAGCAGTGGTGGTTACTACTGGAGCTGGATCCGCCAGCACCCAGGGAAGGGCCTGGAGTGGATTGGGTACATCTATTACAGTGGGAACACCTACTACAACCCGTCCCTCAAGAGTCGAGTTACCATATCAGGAGACACGTCTAAGAACCAGTTCTCCCTGAAGCTGAGGTCTGTGACTGCCGCGGACACGGCCGTGTATTACTGTGCGAGAGGAGGAGCAGCTCGCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCTAGTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGC AGAAGAGCCTCTCCCTGTCTCCGGGTAAA71 Y CAGGTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCACAGACCCTGTCCCTCACCTGCACTGTCTCTGGTGGCTCCATCAGCAGTGGTGGTTTCTACTGGAGCTGGATCCGCCAGCACCCAGGGAAGGGCCTGGAGTGGATTGGGTACATCTATTACAGTGGGAATACCTACTACAACCCGTCCCTCAAGAGTCGAGTTATCATATCAGGAGACACGTCTAAGAACCAGTTCTCCCTGAAGCTGAGCTCTGTGACGGCCGCGGACACGGCCGTGTATTACTGTGCGAGAGGAGGAGCAGCTCGCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCTAGTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGC AGAAGAGCCTCTCCCTGTCTCCGGGTAAA

TABLE 2A Exemplary Anti-hPAC1 Antibody Light Chain Amino Acid SequencesSEQ ID Ab NO: ID LC AA Sequence 2 ADIQMTQSPSSLSASVGDRITITCRASQSISRYLNWYQQKPGKAPKLLIYAASSLQSGIPSRFSGSGSGTDFTLTINSLQPEDFATYFCQQSYSPPFTFGPGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC 2 BDIQMTQSPSSLSASVGDRITITCRASQSISRYLNWYQQKPGKAPKLLIYAASSLQSGIPSRFSGSGSGTDFTLTINSLQPEDFATYFCQQSYSPPFTFGPGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC 4 CDIQLTQSPSFLSASVGDRVTITCRASQSIGRSLHWYQQKPGKAPKLLIKYASQSLSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCHQSSRLPFTFGPGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC 6 DEIVLTQSPATLSLSPGERATLSCRASQSVGRSLHWYQQKPGQAPRLLIKYASQSLSGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCHQSSRLPFTFGPGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT KSFNRGEC 8 EDIQLTQSPSFLSASVGDRVTITCRASQSIGRSLHWYQQKPGKAPKLLIKYASQSLSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCHQSSRLPFTFGPGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC 8 FDIQLTQSPSFLSASVGDRVTITCRASQSIGRSLHWYQQKPGKAPKLLIKYASQSLSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCHQSSRLPFTFGPGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC 8 GDIQLTQSPSFLSASVGDRVTITCRASQSIGRSLHWYQQKPGKAPKLLIKYASQSLSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCHQSSRLPFTFGPGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC 6 HEIVLTQSPATLSLSPGERATLSCRASQSVGRSLHWYQQKPGQAPRLLIKYASQSLSGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCHQSSRLPFTFGPGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT KSFNRGEC 10 JDIQLTQSPSFLSASVGDRVTITCRASQSIGRSLHWYQQKPGKAPKLLFKYASQSLSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCHQSSRLPFTFGPGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC 10 LDIQLTQSPSFLSASVGDRVTITCRASQSIGRSLHWYQQKPGKAPKLLFKYASQSLSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCHQSSRLPFTFGPGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC 12 MEIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSNSGSGTDFTLTISRLEPEDFAVYYCQRYGSSRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC 12 NEIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSNSGSGTDFTLTISRLEPEDFAVYYCQRYGSSRTFGQGTKVEIKRTVAAPSVFIF'PPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC 14 ODIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLLYLGSNRASGVPDRFSGSGSGTDFTLQISRVEAEDVGVYYCMQTLQTPFTFGPGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTH QGLSSPVTKSFNRGEC 16 PDIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLLYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQTLQTPFTFGPGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTH QGLSSPVTKSFNRGEC 16 QDIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLLYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQTLQTPFTFGPGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTH QGLSSPVTKSFNRGEC 18 RKIYTTQSPGTLSLSPGKRATLSCRASQTVSRSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVFYCQQFGSSPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC 20 SDIVMTQSPDSLAVSLGERATIHCKSSQNVLYSSNNKNFLTWYQQKPGQPPKLLIYRASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYFCQQYYSAPFTFGPGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC 22 TDIVMTQSPDSLAVSLGERTTIKCKSSQSVLYRNNNNFLAWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYFCQQYYISPLTFGGGTKVEIKRTVAAPSVF1FPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTH QGLSSPVTKSFNRGEC 24 UDIVMTQSPDSLAVSLGERATINCKSSQSVLYSSNNKHYLAWYRQKPGQPPKLLIYRASTRESGVPDRFSGSGSGTDFTLTISSLQPEDVAVYYCQQYYSSPFTFGPGTKVD1KRTVAAPSVF1FPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC 26 VDIVMTQSPDSLAVSLGERATIHCKSSQSVLYSSNNKNFLTWYQQKPGQPPKLLIYRASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYFCQQYYSAPFTFGPGTRVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC 26 WDIVMTQSPDSLAVSLGERATIHCKSSQSVLYSSNNKNFLTWYQQKPGQPPKLLIYRASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYFCQQYYSAPFTFGPGTRVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRKAKVQWKVDNALQSGNSQESVTEODSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC 28 XDIVMTQSPDSLAVSLGKRATIHCKSSQSVLYSSNNRNFLSWYQQKPGQPPKLLIYRASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYFCQQYYSAPFTFGPGTTVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC 30 YDIVMTQSPDSLAVSLGERATINCKSSOSVLYSSNNKNYLAWYRQKPGQPPKLLIYRASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYHCQQYYSSPFTFGPGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC

TABLE 2B Exemplary Anti-hPAC1 Antibody Heavy Chain Amino AcidS equencesSEQ ID Ab NO: ID HC AA Sequence 32 AQVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSATWNWIRQSPSRGLEWLGRTYYRSKWSNHYAVSVKSRITINPDTSKSQFSLQLNSVTPEDTAVYYCARGTWKQLWFLDHWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYGSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSPGK34 B QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSATWNWIRQSPSRGLEWLGRTYYRSKWSNHYAVSVKSRITINPDTSKSQFSLQLNSVTPEDTAVYYCARGTWKQLWFLDHWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK 36 CQVQLVESGGGVVQPGRSLRLSCAASGFTFSYYAIHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGYDLLTGYPDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK 38 DQVQLVESGGGVVQPGRSLRLSCAASGETFSRFAMHWVRQAPGKGLEWVAVISYDGGNKYYAESVKGRFTISRDNSKNTLYLQMNSLRAEDTALFYCARGYDVLTGYPDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVELEPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYGSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNV FSCSVMHEALHNHYTQKSLSLSPGK 40E QVQLVESGAEVVKPGASVKVSCKASGFTFSRFAMHWVRQAPGQGLEWMGVISYDGGNKYYAESVKGRVTMTRDTSTSTLYMELSSLRSEDTAVYYCARGYDVLTGYPDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYGSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSPGK42 F QVQLVESGAEVVKPGASVKVSCKASGFTFSRFAMHWVRQAPGQGLEWMGVISYDGGNKYYAESVKGRVTMTRDTSTSTLYMELSSLRSEDTAVYYCARGYDVLTGYPDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVELEPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK 44 GQVQLVESGGGVVQPGRSLRLSCAASGETFSRFAMHWVRQAPGKGLEWVAVISYDGGNKYYAESVKGRFTISRDNSKNTLYLQMNSLRAEDTALFYCARGYDVLTGYPDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK 44 HQVQLVESGGGVVQPGRSLRLSCAASGFTFSRFAMHWVRQAPGKGLEWVAVISYDGGNKYYAESVKGRFTISRDNSKNTLYLQMNSLRAEDTALFYCARGYDVLTGYPDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK 46 JQVQLVESGGGVVQPGRSLRLSCAASGFTFSRYAMHWVRQASGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLLMSSLRAEDTAVFYCARGYDILTGYPDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYGSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSPGK 48L QVQLVESGGGVVQPGRSLRLSCAASGFTFSRYAMHWVRQASGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLLMSSLRAEDTAVFYCARGYDILTGYPDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK 50 MQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWINAYNGHTNYAQTFQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARELELRSFYYFGMDVWGQGTTVPVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNV FSCSVMHEALHNHYTQKSLSLSPGK 52N QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWINAYNGHTNYAQTFQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARELELRSFYYFGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNV FSCSVMHEALHNHYTQKSLSLSPGK 54O QVQLVQSGAEVKKSGASLKVSCKASGYIFTRYGVSWVRQAPGQGLEWMGWITTYNGNTNYAQKLQGRVTMTIDTSTSTAYMELRSLRSDDTAVYYCARRVRYSGGYSFDNWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK 56 PQVQLVQSGAEVKKSGASLKVSCKASGYIFTRYGVSWVRQAPGQGLEWMGWITTYNGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARRVRYSGGYSFDNWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNEGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK 58 QQVQLVQSGAEVKKSGASLKVSCKASGYIFTRYGVSWVRQAPGQGLEWMGWITTYNGNTNYAQKLQGRVTMTTDTSTNTAYMELRSLRSDDTAVYYCARRVRYSGGYSFDNWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNEGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLEPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQENSTERVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGK 60 RQVQLQESGPGLVKPSETLSLTCTVSGGSISSYYWSWIRQPAGKGLEWIGRIYTSGSTNYNPSLKSRVTMSIGTSKNQFSLKLSSVTAADTAVYYCAIIASRGWYFDLWGRGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNEGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLEPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTERVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPGK 62 SQVQLQESGPGLVKPSQTLSLTCTVSGGSISSGGYYWSWIRQHPGKGLEWIGYIYYSGNTYYNPSLKSRVTISGDTSKNQFSLKLRSVTAADTAVYYCARGGAARGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNEGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLEPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVM HEALHNHYTQKSLSLSPGK 64 TQVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGRTYYRSRWYNDYAVSVKSRITINPDTSKNQFSLQLNSVTPEDTAVYYCARGVFYSKGAFDIWGQGTMVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK 66 UQVQLQESGPGLVKPSQTLSLTCTVSGGSISRGGYYWSWIRQHPGKGLEWIGYIYYSGNTYYNPSLKSRVIISGDTSKNQLSLKLRSVTAADTAVYYCARGGAARGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVM HEALHNHYTQKSLSLSPGK 68 VQVQLQESGPGLVKPSQTLSLTCTVSGGSISSGGYYWSWIRQHPGKGLEWIGYIYYSGNTYYNPSLKSRVTISGDTSKNQFSLKLRSVTAADTAVYYCTRGGAARGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYGSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPGK 70 WQVQLQESGPGLVKPSQTLSLTCTVSGGSISSGGYYWSWIRQHPGKGLEWIGYIYYSGNTYYNPSLKSRVTISGDTSKNQFSLKLRSVTAADTAVYYCTRGGAARGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVM HEALHNHYTQKSLSLSPGK 62 XQVQLQESGPGLVKPSQTLSLTCTVSGGSISSGGYYWSWIRQHPGKGLEWIGYIYYSGNTYYNPSLKSRVTISGDTSKNQFSLKLRSVTAADTAVYYCARGGAARGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVM HEALHNHYTQKSLSLSPGK 72 YQVQLQESGPGLVKPSQTLSLTCTVSGGSISSGGFYWSWIRQHPGKGLEWIGYIYYSGNTYYNPSLKSRVIISGDTSKNQFSLKLSSVTAADTAVYYCARGGAARGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVM HEALHNHYTQKSLSLSPGKVariable Domains of Antibodies

Also provided are antibodies (and corresponding nucleic acid sequences)that contain an antibody light chain variable region or an antibodyheavy chain variable region, as shown in Tables 4A and 4B below, andimmunologically functional fragments, derivatives, muteins and variantsof these light chain and heavy chain variable regions.

Antibodies of this type can generally be designated by the formula“V_(H)x/V_(L)y,” where “x” corresponds to the number of heavy chainvariable regions and “y” corresponds to the number of the light chainvariable regions.

TABLE 3A  Exemplary Anti-hPAC1 Antibody Light ChainVariable Region Nucleic Acid Sequences SEQ ID Ab NO: IDV_(L) NA Sequence 73 A GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAATCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGGTATTTAAATTGGTATCAACAGAAACCAGGGAAAGCCCCTAAACTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGATCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAACAGTCTGCAACCTGAAGATTTTGCAACTTACTTCTGTCAACAGAGTTACAGTCCCCCATTCACTTTCGGCCCTGGGACCAAAGTGGATA TCAAACGT 73 BGACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAATCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGGTATTTAAATTGGTATCAACAGAAACCAGGGAAAGCCCCTAAACTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGATCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAACAGTCTGCAACCTGAAGATTTTGCAACTTACTTCTGTCAACAGAGTTACAGTCCCCCATTCACTTTCGGCCCTGGGACCAAAGTGGATA TCAAACGT 75 CGATATCCAGCTCACTCAATCGCCATCATTTCTCTCCGCTTCGGTAGGCGACCGGGTCACGATCACATGCAGGGCGTCGCAAAGCATTGGGAGGTCGTTGCATTGGTATCAGCAGAAACCCGGAAAGGCCCCGAAACTTCTGATCAAATACGCATCACAAAGCTTGAGCGGTGTGCCGTCGCGCTTCTCCGGTTCCGGAAGCGGAACGGAATTCACGCTTACAATCTCCTCACTGCAGCCCGAGGATTTCGCGACCTATTACTGTCACCAGTCATCCAGACTCCCGTTTACTTTTGGCCCTGGGACCAAGGTGGACA TTAAGCGT 77 DGAGATCGTACTTACTCAGTCACCCGCCACATTGTCCCTGAGCCCGGGTGAACGGGCGACCCTCAGCTGCCGAGCATCCCAGTCCGTCGGACGATCATTGCACTGGTACCAACAAAAACCGGGCCAGGCCCCCAGACTTCTGATCAAGTATGCGTCACAGAGCTTGTCGGGTATTCCCGCTCGCTTTTCGGGGTCGGGATCCGGGACAGATTTCACGCTCACAATCTCCTCGCTGGAACCCGAGGACTTCGCGGTCTACTATTGTCATCAGTCATCGAGGTTGCCTTTCACGTTTGGACCAGGGACCAAGGTGGACA TTAAGCGT 79 EGATATCCAGCTCACTCAATCGCCATCATTTCTCTCCGCTTCGGTAGGCGACCGGGTCACGATCACATGCAGGGCGTCGCAAAGCATTGGGAGGTCGTTGCATTGGTATCAGCAGAAACCCGGAAAGGCCCCGAAACTTCTGATCAAATACGCATCACAAAGCTTGAGCGGTGTGCCGTCGCGCTTCTCCGGTTCCGGAAGCGGAACGGAATTCACGCTTACAATCTCCTCACTGCAGCCCGAGGATTTCGCGACCTATTACTGTCACCAGTCATCCAGACTCCCGTTTACTTTTGGCCCTGGGACCAAGGTGGACA TTAAGCGT 79 FGATATCCAGCTCACTCAATCGCCATCATTTCTCTCCGCTTCGGTAGGCGACCGGGTCACGATCACATGCAGGGCGTCGCAAAGCATTGGGAGGTCGTTGCATTGGTATCAGCAGAAACCCGGAAAGGCCCCGAAACTTCTGATCAAATACGCATCACAAAGCTTGAGCGGTGTGCCGTCGCGCTTCTCCGGTTCCGGAAGCGGAACGGAATTCACGCTTACAATCTCCTCACTGCAGCCCGAGGATTTCGCGACCTATTACTGTCACCAGTCATCCAGACTCCCGTTTACTTTTGGCCCTGGGACCAAGGTGGACA TTAAGCGT 79 GGATATCCAGCTCACTCAATCGCCATCATTTCTCTCCGCTTCGGTAGGCGACCGGGTCACGATCACATGCAGGGCGTCGCAAAGCATTGGGAGGTCGTTGCATTGGTATCAGCAGAAACCCGGAAAGGCCCCGAAACTTCTGATCAAATACGCATCACAAAGCTTGAGCGGTGTGCCGTCGCGCTTCTCCGGTTCCGGAAGCGGAACGGAATTCACGCTTACAATCTCCTCACTGCAGCCCGAGGATTTCGCGACCTATTACTGTCACCAGTCATCCAGACTCCCGTTTACTTTTGGCCCTGGGACCAAGGTGGACA TTAAGCGT 77 HGAGATCGTACTTACTCAGTCACCCGCCACATTGTCCCTGAGCCCGGGTGAACGGGCGACCCTCAGCTGCCGAGCATCCCAGTCCGTCGGACGATCATTGCACTGGTACCAACAAAAACCGGGCCAGGCCCCCAGACTTCTGATCAAGTATGCGTCACAGAGCTTGTCGGGTATTCCCGCTCGCTTTTCGGGGTCGGGATCCGGGACAGATTTCACGCTCACAATCTCCTCGCTGGAACCCGAGGACTTCGCGGTCTACTATTGTCATCAGTCATCGAGGTTGCCTTTCACGTTTGGACCAGGGACCAAGGTGGACA TTAAGCGT 81 JGATATCCAGCTCACTCAATCGCCATCATTTCTCTCCGCTTCGGTAGGCGACCGGGTCACGATCACATGCAGGGCGTCGCAAAGCATTGGGAGGTCGTTGCATTGGTATCAGCAGAAACCCGGAAAGGCCCCGAAACTTCTGTTCAAATACGCATCACAAAGCTTGAGCGGTGTGCCGTCGCGCTTCTCCGGTTCCGGAAGCGGAACGGAATTCACGCTTACAATCTCCTCACTGCAGCCCGAGGATTTCGCGACCTATTACTGTCACCAGTCATCCAGACTCCCGTTTACTTTTGGCCCTGGGACCAAGGTGGACA TTAAGCGT 81 LGATATCCAGCTCACTCAATCGCCATCATTTCTCTCCGCTTCGGTAGGCGACCGGGTCACGATCACATGCAGGGCGTCGCAAAGCATTGGGAGGTCGTTGCATTGGTATCAGCAGAAACCCGGAAAGGCCCCGAAACTTCTGTTCAAATACGCATCACAAAGCTTGAGCGGTGTGCCGTCGCGCTTCTCCGGTTCCGGAAGCGGAACGGAATTCACGCTTACAATCTCCTCACTGCAGCCCGAGGATTTCGCGACCTATTACTGTCACCAGTCATCCAGACTCCCGTTTACTTTTGGCCCTGGGACCAAGGTGGACA TTAAGCGT 83 MGAAATTGTGTTGACGCAGTCGCCAGGCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTAGCAGCAGCTACTTAGCCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGGTGCATCCAGCAGGGCCACTGGCATCCCAGACAGGTTCAGTAACAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGACTGGAGCCTGAAGATTTTGCAGTGTATTACTGTCAGAGGTATGGTAGCTCACGGACGTTCGGCCAAGGGACCAAGGTGGAAA TCAAACGA 83 NGAAATTGTGTTGACGCAGTCGCCAGGCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTAGCAGCAGCTACTTAGCCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGGTGCATCCAGCAGGGCCACTGGCATCCCAGACAGGTTCAGTAACAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGACTGGAGCCTGAAGATTTTGCAGTGTATTACTGTCAGAGGTATGGTAGCTCACGGACGTTCGGCCAAGGGACCAAGGTGGAAA TCAAACGA 85 OGATATTGTGATGACTCAGTCTCCACTCTCCCTGCCCGTCACCCCTGGAGAGCCGGCCTCCATCTCCTGCAGGTCTAGTCAGAGCCTCCTGCATAGTAATGGATACAACTATTTGGATTGGTACCTGCAGAAGCCAGGGCAGTCTCCACAGCTCCTGCTCTATTTGGGTTCTAATCGGGCCTCCGGGGTCCCTGACAGGTTCAGTGGCAGTGGATCAGGCACAGATTTTACACTGCAAATCAGCAGAGTGGAGGCTGAGGATGTTGGGGTTTATTACTGCATGCAAACTCTACAAACTCCATTCACTTTCGGCCCTGGG ACCAAAGTGGATATCAAACGT 87 PGATATTGTGATGACTCAGTCTCCACTCTCCCTGCCCGTCACCCCTGGAGAGCCGGCCTCCATCTCCTGCAGGTCTAGTCAGAGCCTCCTGCATAGTAATGGATACAACTATTTGGATTGGTACCTGCAGAAGCCAGGGCAGTCTCCACAGCTCCTGCTCTATTTGGGTTCTAATCGGGCCTCCGGGGTCCCTGACAGGTTCAGTGGCAGTGGATCAGGCACAGATTTTACACTGAAAATCAGCAGAGTGGAGGCTGAGGATGTTGGGGTTTATTACTGCATGCAAACTCTACAAACTCCATTCACTTTCGGCCCTGGG ACCAAAGTGGATATCAAACGT 87 QGATATTGTGATGACTCAGTCTCCACTCTCCCTGCCCGTCACCCCTGGAGAGCCGGCCTCCATCTCCTGCAGGTCTAGTCAGAGCCTCCTGCATAGTAATGGATACAACTATTTGGATTGGTACCTGCAGAAGCCAGGGCAGTCTCCACAGCTCCTGCTCTATTTGGGTTCTAATCGGGCCTCCGGGGTCCCTGACAGGTTCAGTGGCAGTGGATCAGGCACAGATTTTACACTGAAAATCAGCAGAGTGGAGGCTGAGGATGTTGGGGTTTATTACTGCATGCAAACTCTACAAACTCCATTCACTTTCGGCCCTGGG ACCAAAGTGGATATCAAACGT 89 RGAAATTGTGTTGACGCAGTCTCCAGGCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGACTGTTAGCAGGAGCTACTTAGCCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGGTGCATCCAGCAGGGCCACTGGCATCCCAGACAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGACTGGAGCCTGAAGATTTTGCCGTGTTTTACTGTCAGCAGTTTGGTAGCTCACCGTGGACGTTCGGCCAAGGGACCAAGGTGG AAATCAAACGT 91 SGACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCACCATCCATTGCAAGTCCAGCCAGAATGTTTTATACAGCTCCAACAATAAGAACTTCTTAACTTGGTACCAGCAGAAACCAGGACAGCCCCCTAAACTGCTCATTTACCGGGCATCTACCCGGGAATCCGGGGTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACGGATTTCACTCTCACTATCAGCAGTCTGCAGGCTGAAGATGTGGCAGTTTATTTCTGTCAGCAATATTATAGTGCTCCATTCACTTTCGGCCCT GGGACCAAAGTGGATATCAAACGT 93 TGACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGACCACCATCAAGTGCAAGTCCAGCCAGAGTGTTTTATACAGATCCAACAATAACAACTTCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTATTGGGCATCTACCCGGGAATCCGGGGTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGGCTGAAGATGTGGCTGTTTATTTCTGTCAGCAATATTATATTTCTCCGCTCACTTTCGGCGGA GGGACCAAGGTGGAGATCAAACGT 95 UGACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTATACAGTTCCAACAATAAGCACTACTTAGCTTGGTACCGGCAGAAACCAGGACAGCCTCCTAAACTGCTCATTTACAGGGCATCTACCCGGGAATCCGGGGTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTGGCAGTGTATTACTGTCAGCAATATTATAGTTCTCCATTCACTTTCGGCC CTGGGACCAAAGTGGATATCAAACGT 97 VGACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCACCATCCACTGCAAGTCCAGCCAGAGTGTTTTATACAGCTCCAACAATAAGAACTTCTTAACTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAACTTCTCATTTACCGGGCATCTACCCGGGAATCCGGGGTTCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGGCTGAAGATGTGGCAGTTTATTTCTGTCAGCAATATTATAGTGCTCCATTCACTTTCGGCCCT GGGACCAGAGTGGATATCAAACGT 97 WGACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCACCATCCACTGCAAGTCCAGCCAGAGTGTTTTATACAGCTCCAACAATAAGAACTTCTTAACTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAACTTCTCATTTACCGGGCATCTACCCGGGAATCCGGGGTTCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGGCTGAAGATGTGGCAGTTTATTTCTGTCAGCAATATTATAGTGCTCCATTCACTTTCGGCCCT GGGACCAGAGTGGATATCAAACGT 99 XGACATCGTGATGACTCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCACCATCCACTGCAAGTCCAGCCAGAGTGTTTTATACAGCTCCAACAATAGGAACTTCTTAAGTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAACTGCTCATTTACCGGGCATCTACCCGGGAATCCGGGGTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGGCTGAAGATGTGGCAGTTTATTTCTGTCAGCAATATTATAGTGCTCCATTCACTTTCGGCCCT GGGACCACAGTGGATATCAAACGT 101 YGACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTATACAGTTCCAACAATAAGAACTACTTAGCTTGGTACCGGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACAGGGCATCTACCCGGGAATCCGGGGTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGGCTGAAGATGTGGCAGTGTATCACTGTCAGCAATATTATAGTTCTCCATTCACTTTCGGCC CTGGGACCAAAGTGGATATCAAACGT

TABLE 3B Exemplary Anti-hPAC1 Antibody Heavy ChainVariable Region Nucleic Acid Sequences SEQ ID Ab NO: IDV_(H) NA Sequence 103 A CAGGTACAGCTGCAGCAGTCAGGTCCAGGACTGGTGAAGCCCTCGCAGACCCTCTCACTCACCTGTGCCATCTCCGGGGACAGTGTCTCTAGCAACAGTGCTACTTGGAACTGGATCAGGCAGTCCCCATCGAGAGGCCTTGAGTGGCTGGGAAGGACATATTACAGGTCCAAGTGGTCTAATCATTATGCAGTATCTGTGAAAAGTCGAATAACCATCAACCCCGACACGTCCAAGAGCCAGTTCTCCCTGCAGCTGAACTCTGTGACTCCCGAGGACACGGCTGTGTATTACTGTGCAAGAGGAACGTGGAAACAGCTATGGTTCCTTGACCACTGGGGCCAGGGAACCCTGGTCACCGTCTCTAGT 103 BCAGGTACAGCTGCAGCAGTCAGGTCCAGGACTGGTGAAGCCCTCGCAGACCCTCTCACTCACCTGTGCCATCTCCGGGGACAGTGTCTCTAGCAACAGTGCTACTTGGAACTGGATCAGGCAGTCCCCATCGAGAGGCCTTGAGTGGCTGGGAAGGACATATTACAGGTCCAAGTGGTCTAATCATTATGCAGTATCTGTGAAAAGTCGAATAACCATCAACCCCGACACGTCCAAGAGCCAGTTCTCCCTGCAGCTGAACTCTGTGACTCCCGAGGACACGGCTGTGTATTACTGTGCAAGAGGAACGTGGAAACAGCTATGGTTCCTTGACCACTGGGGCCAGGGAACCCTGGTCACCGTCTCTAGT 105 CCAGGTGCAGTTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTTACTATGCCATACACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTAGAGTGGGTGGCAGTTATCTCATATGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTGTGTATTACTGTGCGAGAGGATACGATCTTTTGACTGGTTACCCCGACTACTGGGG CCAGGGAACCCTGGTCACCGTCTCCTCA107 D CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGCGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTAGATTTGCCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCATATGATGGAGGAAATAAATACTATGCAGAGTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTCTGTTTTACTGTGCGAGAGGATACGATGTTTTGACTGGTTACCCCGACTACTGGGGC CAGGGAACCCTGGTCACCGTCTCTAGT109 E CAAGTTCAGTTGGTGGAGTCTGGAGCCGAAGTAGTAAAGCCAGGAGCTTCAGTGAAAGTCTCTTGTAAAGCAAGTGGATTCACGTTTAGCCGCTTTGCCATGCATTGGGTGCGGCAAGCTCCCGGTCAGGGGTTGGAGTGGATGGGAGTTATTAGCTATGACGGGGGCAATAAGTACTACGCCGAGTCTGTTAAGGGTCGGGTCACAATGACACGGGACACCTCAACCAGTACACTCTATATGGAACTGTCTAGCCTGAGATCCGAGGACACCGCTGTGTATTATTGCGCTAGGGGGTACGATGTATTGACGGGTTATCCTGATTACTGGGG GCAGGGGACACTCGTAACCGTCTCTAGT109 F CAAGTTCAGTTGGTGGAGTCTGGAGCCGAAGTAGTAAAGCCAGGAGCTTCAGTGAAAGTCTCTTGTAAAGCAAGTGGATTCACGTTTAGCCGCTTTGCCATGCATTGGGTGCGGCAAGCTCCCGGTCAGGGGTTGGAGTGGATGGGAGTTATTAGCTATGACGGGGGCAATAAGTACTACGCCGAGTCTGTTAAGGGTCGGGTCACAATGACACGGGACACCTCAACCAGTACACTCTATATGGAACTGTCTAGCCTGAGATCCGAGGACACCGCTGTGTATTATTGCGCTAGGGGGTACGATGTATTGACGGGTTATCCTGATTACTGGGG GCAGGGGACACTCGTAACCGTCTCTAGT111 G CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGCGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTAGATTTGCCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCATATGATGGAGGAAATAAATACTATGCAGAGTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTCTGTTTTACTGTGCGAGAGGATACGATGTTTTGACTGGTTACCCCGACTACTGGGGC CAGGGAACCCTGGTCACCGTCTCCTCA111 H CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGCGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTAGATTTGCCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCATATGATGGAGGAAATAAATACTATGCAGAGTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTCTGTTTTACTGTGCGAGAGGATACGATGTTTTGACTGGTTACCCCGACTACTGGGGC CAGGGAACCCTGGTCACCGTCTCCTCA113 J CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGCGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTCGCTATGCCATGCACTGGGTCCGCCAGGCTTCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCATATGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTGCTAATGAGCAGCCTGAGAGCTGAGGACACGGCTGTGTTTTACTGTGCGAGAGGATACGATATTTTGACTGGTTACCCCGACTACTGGGGC CAGGGAACCCTGGTCACCGTCTCTAGT115 L CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGCGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTCGCTATGCCATGCACTGGGTCCGCCAGGCTTCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCATATGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTGCTAATGAGCAGCCTGAGAGCTGAGGACACGGCTGTGTTTTACTGTGCGAGAGGATACGATATTTTGACTGGTTACCCCGACTACTGGGGC CAGGGAACCCTGGTCACCGTCTCCTCA117 M CAGGTTCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTCTCCTGCAAGGCTTCTGGTTACACCTTTACCAGCTATGGTATCAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGATGGATCAACGCTTACAATGGTCACACAAACTATGCACAGACGTTCCAGGGCAGAGTCACCATGACCACAGACACATCCACGAGCACAGCCTACATGGAGCTGAGGAGCCTGAGATCTGACGACACGGCCGTGTATTACTGTGCGAGGGAACTGGAACTACGCTCCTTCTATTACTTCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCCCCGTCTCTAGT 119 NCAGGTTCAGCTGGTGCAGTCTGGAGCTGAAGTGAAGAAGCCTGGGGCCTCAGTGAAGGTCTCCTGCAAGGCTTCTGGTTACACCTTTACCAGCTATGGTATCAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGATGGATCAACGCTTACAATGGTCACACAAACTATGCACAGACGTTCCAGGGCAGAGTCACCATGACCACAGACACATCCACGAGCACAGCCTACATGGAACTGAGGAGCCTGAGATCTGACGACACGGCCGTGTATTACTGTGCGAGGGAACTGGAACTACGCTCCTTCTATTACTTCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCTAGT 121 OCAGGTTCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGTCTGGGGCCTCTTTGAAGGTCTCCTGCAAGGCTTCTGGTTACATTTTTACCCGCTATGGTGTCAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGATGGATCACCACTTACAATGGTAACACAAACTATGCACAGAAGCTCCAGGGCAGAGTCACCATGACCATAGACACATCCACGAGCACAGCCTACATGGAACTGAGAAGCCTCAGATCTGACGACACGGCCGTGTATTACTGTGCGAGAAGAGTGCGGTATAGTGGGGGCTACTCGTTTGACAACTGGGGCCAGGGAACCCTGGTCACCGTCTCTAGT 123 PCAGGTTCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGTCTGGGGCCTCTTTGAAGGTCTCCTGCAAGGCTTCTGGTTACATTTTTACCCGCTATGGTGTCAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGATGGATCACCACTTACAATGGTAATACAAACTATGCACAGAAGCTCCAGGGCAGAGTCACCATGACCACAGACACATCCACGAGCACAGCCTACATGGAACTGAGGAGCCTCAGATCTGACGACACGGCCGTGTATTACTGTGCGAGAAGAGTGCGGTACAGTGGGGGCTACTCGTTTGACAACTGGGGCCAGGGAACCCTGGTCACCGTCTCTAGT 125 QCAGGTTCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGTCTGGGGCCTCTTTGAAGGTCTCCTGCAAGGCTTCTGGTTACATTTTTACCCGCTATGGTGTCAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGATGGATCACCACTTACAATGGTAATACAAACTATGCACAGAAACTCCAGGGCAGAGTCACCATGACCACAGACACATCCACGAACACAGCCTACATGGAACTGAGGAGCCTCAGATCTGACGACACGGCCGTGTATTACTGTGCGAGAAGAGTGCGGTATAGTGGGGGCTACTCGTTTGACAACTGGGGCCAGGGAACCCTGGTCACCGTCTCTAGT 127 RCAGGTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCGGAGACCCTGTCCCTCACCTGCACTGTCTCTGGTGGCTCCATCAGTAGTTACTACTGGAGCTGGATCCGGCAGCCCGCCGGGAAGGGACTGGAATGGATTGGGCGTATCTATACCAGTGGGAGCACCAACTACAACCCCTCCCTCAAGAGTCGAGTCACCATGTCAATAGGCACGTCCAAGAACCAGTTCTCCCTGAAGCTGAGCTCTGTGACCGCCGCGGACACGGCCGTGTATTACTGTGCGATTATTGCATCTCGTGGCTGGTACTTCGATCTCTGGGGCCGTGGCA CCCTGGTCACCGTCTCTAGT 129 SCAGGTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCACAGACCCTGTCCCTCACCTGCACTGTCTCTGGTGGCTCCATCAGCAGTGGTGGTTACTACTGGAGCTGGATCCGCCAGCACCCAGGGAAGGGCCTGGAGTGGATTGGGTACATCTATTACAGTGGGAACACCTACTACAACCCGTCCCTCAAGAGTCGAGTTACCATATCAGGAGACACGTCTAAGAACCAGTTCTCCCTGAAGCTGAGGTCTGTGACTGCCGCGGACACGGCCGTGTATTACTGTGCGAGAGGAGGAGCAGCTCGCGGTATGGACGTCTGGGGCCAAG GGACCACGGTCACCGTCTCTAGT 131 TCAGGTACAGCTGCAGCAGTCAGGTCCAGGACTGGTGAAGCCCTCGCAGACCCTCTCACTCACCTGTGCCATCTCCGGGGACAGTGTCTCTAGCAACAGTGCTGCTTGGAACTGGATCAGGCAGTCCCCATCGAGAGGCCTTGAGTGGCTGGGAAGGACATACTACAGGTCCAGGTGGTATAATGATTATGCAGTATCTGTGAAAAGTCGAATAACCATCAACCCAGACACATCCAAGAACCAGTTCTCCCTGCAGCTGAACTCTGTGACTCCCGAGGACACGGCTGTGTATTACTGTGCAAGAGGGGTCTTTTATAGCAAAGGTGCTTTTGATATCTGGGGCCAAGGGACAATGGTCACCGTCTCTAGT 133 U CAGGTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCACAGACCCTGTCCCTCACCTGCACTGTCTCTGGTGGCTCCATCAGCCGTGGTGGTTACTACTGGAGCTGGATCCGCCAGCACCCAGGGAAGGGCCTGGAGTGGATTGGGTACATATATTACAGTGGGAATACCTACTACAACCCGTCCCTCAAGAGTCGAGTTATCATATCAGGAGACACGTCTAAGAACCAGCTCTCCCTGAAGCTGAGGTCTGTGACTGCCGCGGACACGGCCGTGTATTATTGTGCGAGAGGAGGAGCAGCTCGCGGTATGGACGTCTGGGGCCAAG GGACCACGGTCACCGTCTCTAGT 135 VCAGGTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCACAGACCCTGTCCCTCACCTGCACTGTCTCTGGTGGCTCCATCAGCAGTGGTGGTTACTACTGGAGCTGGATCCGCCAGCACCCAGGGAAGGGCCTGGAGTGGATTGGGTACATCTATTACAGTGGGAACACCTACTACAACCCGTCCCTCAAGAGTCGAGTTACCATATCAGGAGACACGTCTAAGAACCAGTTCTCCCTGAAGCTGAGGTCTGTGACTGCCGCGGACACGGCCGTGTATTACTGTACGAGAGGAGGAGCAGCTCGCGGTATGGACGTCTGGGGCCAAG GGACCACGGTCACCGTCTCTAGT 135 WCAGGTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCACAGACCCTGTCCCTCACCTGCACTGTCTCTGGTGGCTCCATCAGCAGTGGTGGTTACTACTGGAGCTGGATCCGCCAGCACCCAGGGAAGGGCCTGGAGTGGATTGGGTACATCTATTACAGTGGGAACACCTACTACAACCCGTCCCTCAAGAGTCGAGTTACCATATCAGGAGACACGTCTAAGAACCAGTTCTCCCTGAAGCTGAGGTCTGTGACTGCCGCGGACACGGCCGTGTATTACTGTACGAGAGGAGGAGCAGCTCGCGGTATGGACGTCTGGGGCCAAG GGACCACGGTCACCGTCTCTAGT 129 XCAGGTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCACAGACCCTGTCCCTCACCTGCACTGTCTCTGGTGGCTCCATCAGCAGTGGTGGTTACTACTGGAGCTGGATCCGCCAGCACCCAGGGAAGGGCCTGGAGTGGATTGGGTACATCTATTACAGTGGGAACACCTACTACAACCCGTCCCTCAAGAGTCGAGTTACCATATCAGGAGACACGTCTAAGAACCAGTTCTCCCTGAAGCTGAGGTCTGTGACTGCCGCGGACACGGCCGTGTATTACTGTGCGAGAGGAGGAGCAGCTCGCGGTATGGACGTCTGGGGCCAAG GGACCACGGTCACCGTCTCTAGT 137 YCAGGTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCACAGACCCTGTCCCTCACCTGCACTGTCTCTGGTGGCTCCATCAGCAGTGGTGGTTTCTACTGGAGCTGGATCCGCCAGCACCCAGGGAAGGGCCTGGAGTGGATTGGGTACATCTATTACAGTGGGAATACCTACTACAACCCGTCCCTCAAGAGTCGAGTTATCATATCAGGAGACACGTCTAAGAACCAGTTCTCCCTGAAGCTGAGCTCTGTGACGGCCGCGGACACGGCCGTGTATTACTGTGCGAGAGGAGGAGCAGCTCGCGGTATGGACGTCTGGGGCCAAG GGACCACGGTCACCGTCTCTAGT

TABLE 4A  Exemplary Anti-hPAC1 Antibody Light ChainVariable Region Amino Acid Sequences SEQ ID Ab NO: ID V_(L) AA Sequence74 A DIQMTQSPSSLSASVGDRITITCRASQSISRYLNWYQQKPGKAPKLLIYAASSLQSGIPSRFSGSGSGTDFTLTINSLQPEDFATYFCQ QSYSPPFTFGPGTKVDIKR 74 BDIQMTQSPSSLSASVGDRITITCRASQSISRYLNWYQQKPGKAPKLLIYAASSLQSGIPSRFSGSGSGTDFTLTINSLQPEDFATYFCQ QSYSPPFTFGPGTKVDIKR 76 CDIQLTQSPSFLSASVGDRVTITCRASQSIGRSLHWYQQKPGKAPKLLIKYASQSLSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCH QSSRLPFTFGPGTKVDIKR 78 DEIVLTQSPATLSLSPGERATLSCRASQSVGRSLHWYQQKPGQAPRLLIKYASQSLSGIPARFSGSGSGTDFTLTISSLEPEDFAVYYC HQSSRLPFTFGPGTKVDIKR 80 EDIQLTQSPSFLSASVGDRVTITCRASQSIGRSLHWYQQKPGKAPKLLIKYASQSLSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCH QSSRLPFTFGPGTKVDIKR 80 FDIQLTQSPSFLSASVGDRVTITCRASQSIGRSLHWYQQKPGKAPKLLIKYASQSLSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCH QSSRLPFTFGPGTKVDIKR 80 GDIQLTQSPSFLSASVGDRVTITCRASQSIGRSLHWYQQKPGKAPKLLIKYASQSLSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCH QSSRLPFTFGPGTKVDIKR 78 HEIVLTQSPATLSLSPGERATLSCRASQSVGRSLHWYQQKPGQAPRLLIKYASQSLSGIPARFSGSGSGTDFTLTISSLEPEDFAVYYC HQSSRLPFTFGPGTKVDIKR 82 JDIQLTQSPSFLSASVGDRVTITCRASQSIGRSLHWYQQKPGKAPKLLFKYASQSLSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCH QSSRLPFTFGPGTKVDIKR 82 LDIQLTQSPSFLSASVGDRVTITCRASQSIGRSLHWYQQKPGKAPKLLFKYASQSLSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCH QSSRLPFTFGPGTKVDIKR 84 MEIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSNSGSGTDFTLTISRLEPEDFAVYYC QRYGSSRTFGQGTKVEIKR 84 NEIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSNSGSGTDFTLTISRLEPEDFAVYYC QRYGSSRTFGQGTKVEIKR 86 ODIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLLYLGSNRASGVPDRFSGSGSGTDFTLQISRVEAEDV GVYYCMQTLQTPFTFGPGTKVDIKR88 P DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLLYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDV GVYYCMQTLQTPFTFGPGTKVDIKR88 Q DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLLYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDV GVYYCMQTLQTPFTFGPGTKVDIKR90 R EIVLTQSPGTLSLSPGERATLSCRASQTVSRSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVFY CQQFGSSPWTFGQGTKVEIKR 92 SDIVMTQSPDSLAVSLGERATIHCKSSQNVLYSSNNKNFLTWYQQKPGQPPKLLIYRASTRESGVPDRFSGSGSGTDFTLTISSLQAED VAVYFCQQYYSAPFTFGPGTKVDIKR94 T DIVMTQSPDSLAVSLGERTTIKCKSSQSVLYRSNNNNFLAWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAE DVAVYFCQQYYISPLTFGGGTKVEIKR96 U DIVMTQSPDSLAVSLGERATINCKSSQSVLYSSNNKHYLAWYRQKPGQPPKLLIYRASTRESGVPDRFSGSGSGTDFTLTISSLQPED VAVYYCQQYYSSPFTFGPGTKVDIKR98 V DIVMTQSPDSLAVSLGERATIHCKSSQSVLYSSNNKNFLTWYQQKPGQPPKLLIYRASTRESGVPDRFSGSGSGTDFTLTISSLQAED VAVYFCQQYYSAPFTFGPGTRVDIKR98 W DIVMTQSPDSLAVSLGERATIHCKSSQSVLYSSNNKNFLTWYQQKPGQPPKLLIYRASTRESGVPDRFSGSGSGTDFTLTISSLQAED VAVYFCQQYYSAPFTFGPGTRVDIKR100 X DIVMTQSPDSLAVSLGERATIHCKSSQSVLYSSNNRNFLSWYQQKPGQPPKLLIYRASTRESGVPDRFSGSGSGTDFTLTISSLQAED VAVYFCQQYYSAPFTFGPGTTVDIKR102 Y DIVMTQSPDSLAVSLGERATINCKSSQSVLYSSNNKNYLAWYRQKPGQPPKLLIYRASTRESGVPDRFSGSGSGTDFTLTISSLQAED VAVYHCQQYYSSPFTFGPGTKVDIKR

TABLE 4B  Exemplary Anti-hPAC1 Antibody Heavy ChainVariable Region Amino Acid Sequences SEQ ID Ab NO: ID V_(H) AA Sequence104 A QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSATWNWIRQSPSRGLEWLGRTYYRSKWSNHYAVSVKSRITINPDTSKSQFSLQLNSVTPEDTAVYYCARGTWKQLWFLDHWGQGTLVTVSS 104 BQVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSATWNWIRQSPSRGLEWLGRTYYRSKWSNHYAVSVKSRITINPDTSKSQFSLQLNSVTPEDTAVYYCARGTWKQLWFLDHWGQGTLVTVSS 106 CQVQLVESGGGVVQPGRSLRLSCAASGFTFSYYAIHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGYDLLTGYPDYWGQGTLVTVSS 108 DQVQLVESGGGVVQPGRSLRLSCAASGFTFSRFAMHWVRQAPGKGLEWVAVISYDGGNKYYAESVKGRFTISRDNSKNTLYLQMNSLRAEDTALFYCARGYDVLTGYPDYWGQGTLVTVSS 110 EQVQLVESGAEVVKPGASVKVSCKASGFTFSRFAMHWVRQAPGQGLEWMGVISYDGGNKYYAESVKGRVTMTRDTSTSTLYMELSSLRSEDTAVYYCARGYDVLTGYPDYWGQGTLVTVSS 110 FQVQLVESGAEVVKPGASVKVSCKASGFTFSRFAMHWVRQAPGQGLEWMGVISYDGGNKYYAESVKGRVTMTRDTSTSTLYMELSSLRSEDTAVYYCARGYDVLTGYPDYWGQGTLVTVSS 112 GQVQLVESGGGVVQPGRSLRLSCAASGFTFSRFAMHWVRQAPG KGLEWVAVISYDGGNKYYAESVKGRFTISRDNSKNTLYLQMN SLRAEDTALFYCARGYDVLTGYPDYWGQGTLVTVSS 112 HQVQLVESGGGVVQPGRSLRLSCAASGFTFSRFAMHWVRQAPG KGLEWVAVISYDGGNKYYAESVKGRFTISRDNSKNTLYLQMN SLRAEDTALFYCARGYDVLTGYPDYWGQGTLVTVSS 114 JQVQLVESGGGVVQPGRSLRLSCAASGFTFSRYAMHWVRQASG KGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLLMSS LRAEDTAVFYCARGYDILTGYPDYWGQGTLVTVSS 116 LQVQLVESGGGWQPGRSLRLSCAASGFTFSRYAMHWVRQASG KGLEWVAVISYDGSNKYYADSVKGRFT1SRDNSKNTLYLLMSS LRAEDTAVFYCARGYDILTGYPDYWGQGTLVTVSS 118 MQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPG QGLEWMGWINAYNGHTNYAQTFQGRVTMTTDTSTSTAYMEL RSLRSDDTAVYYCARELELRSFYYFGMDVWGQGTTVPVSS 120 NQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPG QGLEWMGW1NAYNGHTNYAQTFQGRVTMTTDTSTSTAYMEL RSLRSDDTAVYYCARELELRSFYYFGMDVWGQGTTVTVSS 122 OQVQLVQSGAEVKKSGASLKVSCKASGYIFTRYGVSWVRQAPG QGLEWMGWITTYNGNTNYAQKLQGRVTMTTDTSTSTAYMELR SLRSDDTAVYYCARRVRYSGGYSFDNWGQGTLVTVSS 124 PQVQLVQSGAEVKKSGASLKVSCKASGYIFTRYGVSWVRQAPG QGLEWMGWITTYNGNTNYAQKLQGRVTMTTDTSTSTAYMEL RSLRSDDTAVYYCARRVRYSGGYSFDNWGQGTLVTVSS 126 QOVQLVQSGAEVKKSGASLKVSCKASGY1FTRYGVSWVRQAPG QGLEWMGWITTYNGNTNYAQKLQGRVTMTTDTSTNTAYMEL RSLRSDDTAVYYCARRVRYSGGYSFDNWGQGTLVTVSS 128 RQVQLQESGPGLVKPSETLSLTCTVSGGSISSYYWSWIRQPAGKGLEWIGRIYTSGSTNYNPSLKSRVTMSIGTSKNQFSLKLSSVTAADTAVYYCAIIASRGWYFDLWGRGTLVTVSS 130 SQVQLQESGPGLVKPSQTLSLTCTVSGGSISSGGYYWSWIRQHPGKGLEWIGYIYYSGNTYYNPSLKSRVTISGDTSKNQFSLKLRSVTAADTAVYYCARGGAARGMDVWGQGTTVTVSS 132 TQVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGRTYYRSRWYNDYAVSVKSRITINPDTSKNQFSLQLNSVTPEDTAVYYCARGVFYSKGAFDIWGQGTMVTVSS 134 UQVQLQESGPGLVKPSQTLSLTCTVSGGSISRGGYYWSWIRQHPGKGLEWIGYIYYSGNTYYNPSLKSRVIISGDTSKNQLSLKLRSVTAADTAVYYCARGGAARGMDVWGQGTTVTVSS 136 VQVQLQESGPGLVKPSQTLSLTCTVSGGSISSGGYYWSWIRQHPGKGLEWIGYIYYSGNTYYNPSLKSRVTISGDTSKNQFSLKLRSVTAADTAVYYCTRGGAARGMDVWGQGTTVTVSS 136 WQVQLQESGPGLVKPSQTLSLTCTVSGGSISSGGYYWSWIRQHPGKGLEWIGYIYYSGNTYYNPSLKSRVTISGDTSKNQFSLKLRSVTAADTAVYYCTRGGAARGMDVWGQGTTVTVSS 130 XQVQLQESGPGLVKPSQTLSLTCTVSGGSISSGGYYWSWIRQHPGKGLEWIGYIYYSGNTYYNPSLKSRVTISGDTSKNQFSLKLRSVTAADTAVYYCARGGAARGMDVWGQGTTVTVSS 138 YQVQLQESGPGLVKPSQTLSLTCTVSGGSISSGGFYWSWIRQHPGKGLEWIGYIYYSGNTYYNPSLKSRVIISGDTSKNQFSLKLSSVTAADTAVYYCARGGAARGMDVWGQGTTVTVSSEach of the heavy chain variable regions listed in Table 4B may becombined with any of the light chain variable regions shown in Table 4Ato form an antibody.CDRs

The antibodies disclosed herein are polypeptides into which one or moreCDRs are grafted, inserted and/or joined. An antibody can have 1, 2, 3,4, 5 or 6 CDRs. An antibody thus can have, for example, one heavy chainCDR1 (“CDRH1”), and/or one heavy chain CDR2 (“CDRH2”), and/or one heavychain CDR3 (“CDRH3”), and/or one light chain CDR1 (“CDRL1”), and/or onelight chain CDR2 (“CDRL2”), and/or one light chain CDR3 (“CDRL3”). Someantibodies include both a CDRH3 and a CDRL3. Specific heavy and lightchain CDRs are identified in Tables 4A and 4B, respectively.

Complementarity determining regions (CDRs) and framework regions (FR) ofa given antibody may be identified using the system described by Kabatet al. in Sequences of Proteins of Immunological Interest, 5th Ed., USDept. of Health and Human Services, PHS, NIH, NIH Publication no.91-3242, 1991. Certain antibodies that are disclosed herein comprise oneor more amino acid sequences that are identical or have substantialsequence identity to the amino acid sequences of one or more of the CDRspresented in Table 5A (CDRLs) and Table 5B (CDRHs).

TABLE 5A  Exemplary Anti-hPAC1 Antibody Light Chain CDR AminoAcid Sequences SEQ SEQ SEQ Ab ID Unique ID Unique ID Unique ID LC CDR1NO: ID LC CDR2 NO: ID LC CDR3 NO: ID A RASQSISR 141 CDRL1-3 AASSLQ  152CDRL2- QQSYSPPF 158 CDRL3- YLN S 2 T 2 B RASQSISR 141 CDRL1-3 AASSLQ 152 CDRL2- QQSYSPPF 158 CDRL3- YLN S 2 T 2 C RASQSIGR 139 CDRL1-1YASQSL  151 CDRL2- HQSSRLPF 157 CDRL3- SLH S 1 T 1 D RASQSVG 140 CDRL1-2YASQSL  151 CDRL2- HQSSRLPF 157 CDRL3- RSLH S 1 T 1 E RASQSIGR 139CDRL1-1 YASQSL  151 CDRL2- HQSSRLPF 157 CDRL3- SLH S 1 T 1 F RASQSIGR139 CDRL1-1 YASQSL  151 CDRL2- HQSSRLPF 157 CDRL3- SLH S 1 T 1 GRASQSIGR 139 CDRL1-1 YASQSL  151 CDRL2- HQSSRLPF 157 CDRL3- SLH S 1 T 1H RASQSVG 140 CDRL1-2 YASQSL  151 CDRL2- HQSSRLPF 157 CDRL3- RSLH S 1 T1 J RASQSIGR 139 CDRL1-1 YASQSL  151 CDRL2- HQSSRLPF 157 CDRL3- SLHS S 1T 1 L RASQSIGR 139 CDRL1-1 YASQSL  151 CDRL2- HQSSRLPF 157 CDRL3- SLH S1 T 1 M RASQSVSS 142 CDRL1-4 GASSR  153 CDRL2- QRYGSSR 159 CDRL3- SYLAAT 3 T 3 N RASQSVSS 142 CDRL1-4 GASSR 153 CDRL2- QRYGSSR 159 CDRL3- SYLAAT 3 T 3 O RSSQSLLH 144 CDRL1-6 LGSNR 154 CDRL2- MQTLQTP 161 CDRL3-SNGYNYL AS 4 FT 5 D P RSSQSLLH 144 CDRL1-6 LGSNR 154 CDRL2- MQTLQTP 161CDRL3- SNGYNYL AS 4 FT 5 D Q RSSQSLLH 144 CDRL1-6 LGSNR 154 CDRL2-MQTLQTP 161 CDRL3- SNGYNYL AS 4 FT 5 D R RASQTVS 143 CDRL1-5 GASSR 153CDRL2- QQFGSSP 160 CDRL3- RSYLA AT 3 WT 4 S KSSQNVL 145 CDRL1-7 RASTRE155 CDRL2- QQYYSAP 162 CDRL3- YSSNNKN S 5 FT 6 FLT T KSSQSVL 146 CDRL1-8WASTR 156 CDRL2- QQYYISPL 163 CDRL3- YRSNNNN ES 6 T 7 FLA U KSSQSVL 147CDRL1-9 RASTRE 155 CDRL2- QQYYSSPF 164 CDRL3- YSSNNKH S 5 T 8 YLA VKSSQSVL 148 CDRL1- RASTRE 155 CDRL2- QQYYSAP 162 CDRL3- YSSNNKN 10 S 5FT 6 FLT W KSSQSVL 148 CDRL1- RASTRE 155 CDRL2- QQYYSAP 162 CDRL3-YSSNNKN 10 S 5 FT 6 FLT X KSSQSVL 150 CDRL1- RASTRE 155 CDRL2- QQYYSAP162 CDRL3- YSSNNRN 12 S 5 FT 6 FLS Y KSSQSVL 149 CDRL1- RASTRE 155CDRL2- QQYYSSPF 164 CDRL3- YSSNNKN 11 S 5 T 8 YLA

TABLE 5B Exemplary Anti-hPAC1 Antibody Heavy Chain CDR Amino Acid Sequences SEQSEQ SEQ Ab HC ID Unique ID Unique HC ID Unique ID CDR1 NO: ID HC CDR2NO: ID CDR3 NO: ID A SNSAT 172 CDRH1-8 RTYYRSKWSNH 177 CDRH2-2 GTWKQL186 CDRH3- WN YAVSVKS WFLDH 3 B SNSAT 172 CDRH1-8 RTYYRSKWSNH 177CDRH2-2 GTWKQL 186 CDRH3- WN YAVSVKS WFLDH 3 C YYAIH 175 CDRH1-11VISYDGSNKYYA 180 CDRH2-5 GYDLLT 192 CDRH3- DSVKG GYPDY 6 D RFAMH 165CDRH1-1 VISYDGGNKYY 179 CDRH2-4 GYDVLT 189 CDRH3- AESVKG GYPDY 6 E RFAMH165 CDRH1-1 VISYDGGNKYY 179 CDRH2-4 GYDVLT 189 CDRH3- AESVKG GYPDY 6 FRFAMH 165 CDRH1-1 VISYDGGNKYY 179 CDRH2-4 GYDVLT 189 CDRH3- AESVKG GYPDY6 G RFAMH 165 CDRH1-1 VISYDGGNKYY 179 CDRH2-4 GYDVLT 189 CDRH3- AESVKGGYPDY 6 H RFAMH 165 CDRH1-1 VISYDGGNKYY 179 CDRH2-4 GYDVLT 189 CDRH3-AESVKG GYPDY 6 J RYAMH 167 CDRH1-3 VISYDGSNKYYA 180 CDRH2-5 GYDILT 188CDRH3- DSVKG GYPDY 5 L RYAMH 167 CDRH1-3 VISYDGSNKYYA 180 CDRH2-5 GYDILT188 CDRH3- DSVKG GYPDY 5 M SYGIS 173 CDRH1-9 WINAYNGHTNY 181 CDRH2-6ELELRSF 184 CDRH3- AQTFQG YYFGMDV 1 N SYGIS 173 CDRH1-9 WINAYNGHTNY 181CDRH2-6 ELELRSF 184 CDRH3- AQTFQG YYFGMDV 1 O RYGVS 168 CDRH1-4WITTYNGNTNY 182 CDRH2-7 RVRYSG 191 CDRH3- AQKLQG GYSFDN 8 P RYGVS 168CDRH1-4 WITTYNGNTNY 182 CDRH2-7 RVRYSG 191 CDRH3- AQKLQG GYSFDN 8 QRYGVS 168 CDRH1-4 WITTYNGNTNY 182 CDRH2-7 RVRYSG 191 CDRH3- AQKLQGGYSFDN 8 R SYYWS 174 CDRH1-10 RIYTSGSTNYNP 176 CDRH2-1 IASRGW 190 CDRH3-SLKS YFDL 7 S SGGYY 170 CDRH1-6 YIYYSGNTYYNP 183 CDRH2-8 GGAARG 185CDRH3- WS SLKS MDV 2 T SNSAA 171 CDRH1-7 RTYYRSRWYND 178 CDRH2-3 GVFYSK187 CDRH3- WN YAVSVKS GAFDI 4 U RGGYY 166 CDRH1-2 YIYYSGNTYYNP 183CDRH2-8 GGAARG 185 CDRH3- WS SLKS MDV 2 V SGGYY 170 CDRH1-6 YIYYSGNTYYNP183 CDRH2-8 GGAARG 185 CDRH3- WS SLKS MDV 2 W SGGYY 170 CDRH1-6YIYYSGNTYYNP 183 CDRH2-8 GGAARG 185 CDRH3- WS SLKS MDV 2 X SGGYY 170CDRH1-6 YIYYSGNTYYNP 183 CDRH2-8 GGAARG 185 CDRH3- WS SLKS MDV 2 Y SGGFY169 CDRH1-5 YIYYSGNTYYNP 183 CDRH2-8 GGAARG 185 CDRH3- WS SLKS MDV 2

TABLE 5C SEQ ID NOs of CDR Sequences SEQ ID NO: Unique ID Sequence 139CDRL1-1 RASQSIGRSLH 140 CDRL1-2 RASQSVGRSLH 141 CDRL1-3 RASQSISRYLN 142CDRL1-4 RASQSVSSSYLA 143 CDRL1-5 RASQTVSRSYLA 144 CDRL1-6RSSQSLLHSNGYNYLD 145 CDRL1-7 KSSQNVLYSSNNKNFLT 146 CDRL1-8KSSQSVLYRSNNNNFLA 147 CDRL1-9 KSSQSVLYSSNNKHYLA 148 CDRL1-10KSSQSVLYSSNNKNFLT 149 CDRL1-11 KSSQSVLYSSNNKNYLA 150 CDRL1-12KSSQSVLYSSNNRNFLS 151 CDRL2-1 YASQSLS 152 CDRL2-2 AASSLQS 153 CDRL2-3GASSRAT 154 CDRL2-4 LGSNRAS 155 CDRL2-5 RASTRES 156 CDRL2-6 WASTRES 157CDRL3-1 HQSSRLPFT 158 CDRL3-2 QQSYSPPFT 159 CDRL3-3 QRYGSSRT 160 CDRL3-4QQFGSSPWT 161 CDRL3-5 MQTLQTPFT 162 CDRL3-6 QQYYSAPFT 163 CDRL3-7QQYYISPLT 164 CDRL3-8 QQYYSSPFT 165 CDRH1-1 RFAMH 166 CDRH1-2 RGGYYWS167 CDRH1-3 RYAMH 168 CDRH1-4 RYGVS 169 CDRH1-5 SGGFYWS 170 CDRH1-6SGGYYWS 171 CDRH1-7 SNSAAWN 172 CDRH1-8 SNSATWN 173 CDRH1-9 SYGIS 174CDRH1-10 SYYWS 175 CDRH1-11 YYAIH 176 CDRH2-1 RIYTSGSTNYNPSLKS 177CDRH2-2 RTYYRSKWSNHYAVSVKS 178 CDRH2-3 RTYYRSRWYNDYAVSVKS 179 CDRH2-4VISYDGGNKYYAESVKG 180 CDRH2-5 VISYDGSNKYYADSVKG 181 CDRH2-6WINAYNGHTNYAQTFQG 182 CDRH2-7 WITTYNGNTNYAQKLQG 183 CDRH2-8YIYYSGNTYYNPSLKS 184 CDRH3-1 ELELRSFYYFGMDV 185 CDRH3-2 GGAARGMDV 186CDRH3-3 GTWKQLWFLDH 187 CDRH3-4 GVFYSKGAFDI 188 CDRH3-5 GYDILTGYPDY 189CDRH3-6 GYDVLTGYPDY 190 CDRH3-7 IASRGWYFDL 191 CDRH3-8 RVRYSGGYSFDN

The structure and properties of CDRs within a naturally occurringantibody has been described, supra. Briefly, in a traditional antibody,the CDRs are embedded within a framework in the heavy and light chainvariable region where they constitute the regions responsible forantigen binding and recognition. A variable region comprises at leastthree heavy or light chain CDRs, see, supra (Kabat et al., 1991,Sequences of Proteins of Immunological Interest, Public Health ServiceN.I.H., Bethesda, Md.; see also Chothia and Lesk, 1987, J. Mol. Biol.196:901-917; Chothia et al., 1989, Nature 342: 877-883), within aframework region (designated framework regions 1-4, FR1, FR2, FR3, andFR4, by Kabat et al., 1991, supra; see also Chothia and Lesk, 1987,supra). The CDRs provided herein, however, may not only be used todefine the antigen binding domain of a traditional antibody structure,but may be embedded in a variety of other polypeptide structures, asdescribed herein.

Some of the antibodies disclosed herein share certain regions orsequences with other antibodies disclosed herein. These relationshipsare summarized in Table 6, below.

TABLE 6 Antibody types and shared sequences LC Full HC Full LC Var HCVar Ab ID IgG type same same same same A IgG1 A, B A, B A, B B IgG2 A, BA, B A, B C IgG2 D IgG1 D, H D, H D, G, H E IgG1 E, F, G E, F, G E, F FIgG2 E, F, G E, F, G E, F G IgG2 E, F, G G, H E, F, G D, G, H H IgG2 D,H G, H D, H D, G, H J IgG1 J, L J, L J, L L IgG2 J, L J, L J, L M IgG2M, N M, N N IgG2 M, N M, N O IgG2 P IgG2 P, Q P, Q Q IgG2 P, Q P, Q RIgG2 S IgG2 S, X S, X T IgG2 U IgG2 V IgG1 V, W V, W V, W W IgG2 V, W V,W V, W X IgG2 S, X S, X Y IgG2

In one aspect, the isolated antibodies provided herein can be amonoclonal antibody, a polyclonal antibody, a recombinant antibody, ahuman antibody, a humanized antibody, a chimeric antibody, amultispecific antibody, or an antibody antigen binding fragment thereof.

In another embodiment, the antibody fragment of the isolated antibodiesprovided herein can be a Fab fragment, a Fab′ fragment, an F(ab′)₂fragment, an Fv fragment, a diabody, or a single chain antibodymolecule.

In a further embodiment, the isolated antibody provided herein is ahuman antibody and can be of the IgG1-, IgG2-IgG3- or IgG4-type. In afurther embodiment, the isolated antibody is an IgG1- or IgG2-type. In afurther embodiment, the isolated antibody is an IgG2-type. In a furtherembodiment, the isolated antibody is an IgG1-type. In a furtherembodiment, the IgG1-type antibody is an aglycosylated IgG1 antibody. Ina further embodiment, the aglycosylated IgG1 antibody has an N297Gmutation.

In another embodiment, the antibody consists of a just a light or aheavy chain polypeptide as set forth in Tables 2A-2B. In someembodiments, the antibody consists just of a light chain variable orheavy chain variable domain such as those listed in Tables 4A-4B. Suchantibodies can be pegylated with one or more PEG molecules.

In yet another aspect, the isolated antibody provided herein can becoupled to a labeling group and can compete for binding to theextracellular portion of human PAC1 with an antibody of one of theisolated antibodies provided herein. In one embodiment, the isolatedantibody provided herein can reduce monocyte chemotaxis, inhibitmonocyte migration into tumors or inhibit accumulation and function oftumor associated macrophage in a tumor when administered to a patient.

As will be appreciated by those in the art, for any antibody with morethan one CDR from the depicted sequences, any combination of CDRsindependently selected from the depicted sequences is useful. Thus,antibodies with one, two, three, four, five or six of independentlyselected CDRs can be generated. However, as will be appreciated by thosein the art, specific embodiments generally utilize combinations of CDRsthat are non-repetitive, e.g., antibodies are generally not made withtwo CDRH2 regions, etc.

Monoclonal Antibodies

The antibodies that are provided include monoclonal antibodies that bindto PAC1. Monoclonal antibodies may be produced using any technique knownin the art, e.g., by immortalizing spleen cells harvested from thetransgenic animal after completion of the immunization schedule. Thespleen cells can be immortalized using any technique known in the art,e.g., by fusing them with myeloma cells to produce hybridomas. Myelomacells for use in hybridoma-producing fusion procedures preferably arenon-antibody-producing, have high fusion efficiency, and enzymedeficiencies that render them incapable of growing in certain selectivemedia which support the growth of only the desired fused cells(hybridomas). Examples of suitable cell lines for use in mouse fusionsinclude Sp-20, P3-X63/Ag8, P3-X63-Ag8.653, NS1/1.Ag 4 1, Sp210-Ag14, FO,NSO/U, MPC-11, MPC11-X45-GTG 1.7 and S194/5XXO Bul; examples of celllines used in rat fusions include R210.RCY3, Y3-Ag 1.2.3, IR983F and4B210. Other cell lines useful for cell fusions are U-266, GM1500-GRG2,LICR-LON-HMy2 and UC729-6.

In some instances, a hybridoma cell line is produced by immunizing ananimal (e.g., a transgenic animal having human immunoglobulin sequences)with a PAC1 immunogen; harvesting spleen cells from the immunizedanimal; fusing the harvested spleen cells to a myeloma cell line,thereby generating hybridoma cells; establishing hybridoma cell linesfrom the hybridoma cells, and identifying a hybridoma cell line thatproduces an antibody that binds PAC1 (e.g., as described in Examples1-3, below). Such hybridoma cell lines, and anti-PAC1 monoclonalantibodies produced by them, are aspects of the present application.

Monoclonal antibodies secreted by a hybridoma cell line can be purifiedusing any technique known in the art.

Chimeric and Humanized Antibodies

Chimeric and humanized antibodies based upon the foregoing sequences arealso provided. Monoclonal antibodies for use as therapeutic agents maybe modified in various ways prior to use. One example is a chimericantibody, which is an antibody composed of protein segments fromdifferent antibodies that are covalently joined to produce functionalimmunoglobulin light or heavy chains or immunologically functionalportions thereof. Generally, a portion of the heavy chain and/or lightchain is identical with or homologous to a corresponding sequence inantibodies derived from a particular species or belonging to aparticular antibody class or subclass, while the remainder of thechain(s) is/are identical with or homologous to a corresponding sequencein antibodies derived from another species or belonging to anotherantibody class or subclass. For methods relating to chimeric antibodies,see, for example, U.S. Pat. No. 4,816,567; and Morrison et al., 1985,Proc. Natl. Acad. Sci. USA 81:6851-6855, which are hereby incorporatedby reference. CDR grafting is described, for example, in U.S. Pat. Nos.6,180,370, 5,693,762, 5,693,761, 5,585,089, and 5,530,101.

Generally, the goal of making a chimeric antibody is to create a chimerain which the number of amino acids from the intended patient species ismaximized. One example is the “CDR-grafted” antibody, in which theantibody comprises one or more complementarity determining regions(CDRs) from a particular species or belonging to a particular antibodyclass or subclass, while the remainder of the antibody chain(s) is/areidentical with or homologous to a corresponding sequence in antibodiesderived from another species or belonging to another antibody class orsubclass. For use in humans, the variable region or selected CDRs from arodent antibody often are grafted into a human antibody, replacing thenaturally-occurring variable regions or CDRs of the human antibody.

One useful type of chimeric antibody is a “humanized” antibody.Generally, a humanized antibody is produced from a monoclonal antibodyraised initially in a non-human animal. Certain amino acid residues inthis monoclonal antibody, typically from non-antigen recognizingportions of the antibody, are modified to be homologous to correspondingresidues in a human antibody of corresponding isotype. Humanization canbe performed, for example, using various methods by substituting atleast a portion of a rodent variable region for the correspondingregions of a human antibody (see, e.g., U.S. Pat. Nos. 5,585,089, and5,693,762; Jones et al., 1986, Nature 321:522-525; Riechmann et al.,1988, Nature 332:323-27; Verhoeyen et al., 1988, Science 239:1534-1536),

In one aspect, the CDRs of the light and heavy chain variable regions ofthe antibodies provided herein are grafted to framework regions (FRs)from antibodies from the same, or a different, phylogenetic species. Forexample, the CDRs of the heavy and light chain variable regionsdescribed herein can be grafted to consensus human FRs. To createconsensus human FRs, FRs from several human heavy chain or light chainamino acid sequences may be aligned to identify a consensus amino acidsequence. In other embodiments, the FRs of a heavy chain or light chaindisclosed herein are replaced with the FRs from a different heavy chainor light chain. In one aspect, rare amino acids in the FRs of the heavyand light chains of anti-PAC1 antibody are not replaced, while the restof the FR amino acids are replaced. A “rare amino acid” is a specificamino acid that is in a position in which this particular amino acid isnot usually found in an FR. Alternatively, the grafted variable regionsfrom the one heavy or light chain may be used with a constant regionthat is different from the constant region of that particular heavy orlight chain as disclosed herein. In other embodiments, the graftedvariable regions are part of a single chain Fv antibody.

In certain embodiments, constant regions from species other than humancan be used along with the human variable region(s) to produce hybridantibodies.

Fully Human Antibodies

Fully human antibodies are also provided. Methods are available formaking fully human antibodies specific for a given antigen withoutexposing human beings to the antigen (“fully human antibodies”). Onespecific means provided for implementing the production of fully humanantibodies is the “humanization” of the mouse humoral immune system.Introduction of human immunoglobulin (Ig) loci into mice in which theendogenous Ig genes have been inactivated is one means of producingfully human monoclonal antibodies (mAbs) in mouse, an animal that can beimmunized with any desirable antigen. Using fully human antibodies canminimize the immunogenic and allergic responses that can sometimes becaused by administering mouse or mouse-derived mAbs to humans astherapeutic agents.

Fully human antibodies can be produced by immunizing transgenic animals(usually mice) that are capable of producing a repertoire of humanantibodies in the absence of endogenous immunoglobulin production.Antigens for this purpose typically have six or more contiguous aminoacids, and optionally are conjugated to a carrier, such as a hapten.See, e.g., Jakobovits et al., 1993, Proc. Natl. Acad. Sci. USA90:2551-2555; Jakobovits et al., 1993, Nature 362:255-258; andBruggermann et al., 1993, Year in Immunol. 7:33. In one example of sucha method, transgenic animals are produced by incapacitating theendogenous mouse immunoglobulin loci encoding the mouse heavy and lightimmunoglobulin chains therein, and inserting into the mouse genome largefragments of human genome DNA containing loci that encode human heavyand light chain proteins. Partially modified animals, which have lessthan the full complement of human immunoglobulin loci, are thencross-bred to obtain an animal having all of the desired immune systemmodifications. When administered an immunogen, these transgenic animalsproduce antibodies that are immunospecific for the immunogen but havehuman rather than murine amino acid sequences, including the variableregions. For further details of such methods, see, for example,WO96/33735 and WO94/02602. Additional methods relating to transgenicmice for making human antibodies are described in U.S. Pat. Nos.5,545,807; 6,713,610; 6,673,986; 6,162,963; 5,545,807; 6,300,129;6,255,458; 5,877,397; 5,874,299 and 5,545,806; in PCT publicationsWO91/10741, WO90/04036, and in EP 546073B1 and EP 546073A1.

The transgenic mice described above, referred to herein as “HuMab” mice,contain a human immunoglobulin gene minilocus that encodes unrearrangedhuman heavy ([mu] and [gamma]) and [kappa] light chain immunoglobulinsequences, together with targeted mutations that inactivate theendogenous [mu] and [kappa] chain loci (Lonberg et al., 1994, Nature368:856-859). Accordingly, the mice exhibit reduced expression of mouseIgM or [kappa] and in response to immunization, and the introduced humanheavy and light chain transgenes undergo class switching and somaticmutation to generate high affinity human IgG [kappa] monoclonalantibodies (Lonberg et al., supra.; Lonberg and Huszar, 1995, Intern.Rev. Immunol. 13: 65-93; Harding and Lonberg, 1995, Ann. N.Y Acad. Sci.764:536-546). The preparation of HuMab mice is described in detail inTaylor et al., 1992, Nucleic Acids Research 20:6287-6295; Chen et al.,1993, International Immunology 5:647-656; Tuaillon et al., 1994, J.Immunol. 152:2912-2920; Lonberg et al., 1994, Nature 368:856-859;Lonberg, 1994, Handbook of Exp. Pharmacology 113:49-101; Taylor et al.,1994, International Immunology 6:579-591; Lonberg and Huszar, 1995,Intern. Rev. Immunol. 13:65-93; Harding and Lonberg, 1995, Ann. N.YAcad. Sci. 764:536-546; Fishwild et al., 1996, Nature Biotechnology14:845-851; the foregoing references are hereby incorporated byreference in their entirety for all purposes. See, further U.S. Pat.Nos. 5,545,806; 5,569,825; 5,625,126; 5,633,425; 5,789,650; 5,877,397;5,661,016; 5,814,318; 5,874,299; and 5,770,429; as well as U.S. Pat. No.5,545,807; International Publication Nos. WO 93/1227; WO 92/22646; andWO 92/03918, the disclosures of all of which are hereby incorporated byreference in their entirety for all purposes. Technologies utilized forproducing human antibodies in these transgenic mice are disclosed alsoin WO 98/24893, and Mendez et al., 1997, Nature Genetics 15:146-156,which are hereby incorporated by reference. For example, the HCo7 andHCo12 transgenic mice strains can be used to generate anti-PAC1antibodies. Further details regarding the production of human antibodiesusing transgenic mice are provided in the examples below.

Using hybridoma technology, antigen-specific human mAbs with the desiredspecificity can be produced and selected from the transgenic mice suchas those described above. Such antibodies may be cloned and expressedusing a suitable vector and host cell, or the antibodies can beharvested from cultured hybridoma cells.

Fully human antibodies can also be derived from phage-display libraries(as disclosed in Hoogenboom et al., 1991, J. Mol. Biol. 227:381; andMarks et al., 1991, J. Mol. Biol. 222:581). Phage display techniquesmimic immune selection through the display of antibody repertoires onthe surface of filamentous bacteriophage, and subsequent selection ofphage by their binding to an antigen of choice. One such technique isdescribed in PCT Publication No. WO 99/10494 (hereby incorporated byreference), which describes the isolation of high affinity andfunctional agonistic antibodies for MPL- and msk-receptors using such anapproach.

Bispecific or Bifunctional Antibodies

The antibodies that are provided also include bispecific andbifunctional antibodies that include one or more CDRs or one or morevariable regions as described above. A bispecific or bifunctionalantibody in some instances is an artificial hybrid antibody having twodifferent heavy/light chain pairs and two different binding sites.Bispecific antibodies may be produced by a variety of methods including,but not limited to, fusion of hybridomas or linking of Fab′ fragments.See, e.g., Songsivilai and Lachmann, 1990, Clin. Exp. Immunol.79:315-321; Kostelny et al., 1992, J. Immunol. 148:1547-1553.

Various Other Forms

Some of the antibodies that are provided are variant forms of theantibodies disclosed above. For instance, some of the antibodies haveone or more conservative amino acid substitutions in one or more of theheavy or light chains, variable regions or CDRs listed above.

Naturally-occurring amino acids may be divided into classes based oncommon side chain properties:

1) hydrophobic: norleucine, Met, Ala, Val, Leu, Ile;

2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gln;

3) acidic: Asp, Glu;

4) basic: His, Lys, Arg;

5) residues that influence chain orientation: Gly, Pro; and

6) aromatic: Trp, Tyr, Phe.

Conservative amino acid substitutions may involve exchange of a memberof one of these classes with another member of the same class.Conservative amino acid substitutions may encompass non-naturallyoccurring amino acid residues, which are typically incorporated bychemical peptide synthesis rather than by synthesis in biologicalsystems. These include peptidomimetics and other reversed or invertedforms of amino acid moieties.

Non-conservative substitutions may involve the exchange of a member ofone of the above classes for a member from another class. Suchsubstituted residues may be introduced into regions of the antibody thatare homologous with human antibodies, or into the non-homologous regionsof the molecule.

In making such changes, according to certain embodiments, thehydropathic index of amino acids may be considered. The hydropathicprofile of a protein is calculated by assigning each amino acid anumerical value (“hydropathy index”) and then repetitively averagingthese values along the peptide chain. Each amino acid has been assigneda hydropathic index on the basis of its hydrophobicity and chargecharacteristics. They are: isoleucine (+4.5); valine (+4.2); leucine(+3.8); phenylalanine (+2.8); cysteine/cystine (+2.5); methionine(+1.9); alanine (+1.8); glycine (−0.4); threonine (−0.7); serine (−0.8);tryptophan (−0.9); tyrosine (−1.3); proline (−1.6); histidine (−3.2);glutamate (−3.5); glutamine (−3.5); aspartate (−3.5); asparagine (−3.5);lysine (−3.9); and arginine (−4.5).

The importance of the hydropathic profile in conferring interactivebiological function on a protein is understood in the art (see, e.g.,Kyte et al., 1982, J. Mol. Biol. 157:105-131). It is known that certainamino acids may be substituted for other amino acids having a similarhydropathic index or score and still retain a similar biologicalactivity. In making changes based upon the hydropathic index, in certainembodiments, the substitution of amino acids whose hydropathic indicesare within ±2 is included. In some aspects, those which are within ±1are included, and in other aspects, those within ±0.5 are included.

It is also understood in the art that the substitution of like aminoacids can be made effectively on the basis of hydrophilicity,particularly where the biologically functional protein or peptidethereby created is intended for use in immunological embodiments, as inthe present case. In certain embodiments, the greatest local averagehydrophilicity of a protein, as governed by the hydrophilicity of itsadjacent amino acids, correlates with its immunogenicity andantigen-binding or immunogenicity, that is, with a biological propertyof the protein.

The following hydrophilicity values have been assigned to these aminoacid residues: arginine (+3.0); lysine (+3.0); aspartate (+3.0±1);glutamate (+3.0±1); serine (+0.3); asparagine (+0.2); glutamine (+0.2);glycine (0); threonine (−0.4); proline (−0.5±1); alanine (−0.5);histidine (−0.5); cysteine (−1.0); methionine (−1.3); valine (−1.5);leucine (−1.8); isoleucine (−1.8); tyrosine (−2.3); phenylalanine (−2.5)and tryptophan (−3.4). In making changes based upon similarhydrophilicity values, in certain embodiments, the substitution of aminoacids whose hydrophilicity values are within ±2 is included, in otherembodiments, those which are within ±1 are included, and in still otherembodiments, those within ±0.5 are included. In some instances, one mayalso identify epitopes from primary amino acid sequences on the basis ofhydrophilicity. These regions are also referred to as “epitopic coreregions.”

Exemplary conservative amino acid substitutions are set forth in Table7.

TABLE 7 Conservative Amino Acid Substitutions Original Residue ExemplarySubstitutions Ala Ser Arg Lys Asn Gln, His Asp Glu Cys Ser Gln Asn GluAsp Gly Pro His Asn, Gln Ile Leu, Val Leu Ile, Val Lys Arg, Gln, Glu MetLeu, Ile Phe Met, Leu, Tyr Ser Thr Thr Ser Trp Tyr Tyr Trp, Phe Val Ile,Leu

A skilled artisan will be able to determine suitable variants ofpolypeptides as set forth herein using well-known techniques. Oneskilled in the art may identify suitable areas of the molecule that maybe changed without destroying activity by targeting regions not believedto be important for activity. The skilled artisan also will be able toidentify residues and portions of the molecules that are conserved amongsimilar polypeptides. In further embodiments, even areas that may beimportant for biological activity or for structure may be subject toconservative amino acid substitutions without destroying the biologicalactivity or without adversely affecting the polypeptide structure.

Additionally, one skilled in the art can review structure-functionstudies identifying residues in similar polypeptides that are importantfor activity or structure. In view of such a comparison, one can predictthe importance of amino acid residues in a protein that correspond toamino acid residues important for activity or structure in similarproteins. One skilled in the art may opt for chemically similar aminoacid substitutions for such predicted important amino acid residues.

One skilled in the art can also analyze the 3-dimensional structure andamino acid sequence in relation to that structure in similarpolypeptides. In view of such information, one skilled in the art maypredict the alignment of amino acid residues of an antibody with respectto its three dimensional structure. One skilled in the art may choosenot to make radical changes to amino acid residues predicted to be onthe surface of the protein, since such residues may be involved inimportant interactions with other molecules. Moreover, one skilled inthe art may generate test variants containing a single amino acidsubstitution at each desired amino acid residue. These variants can thenbe screened using assays for PAC1 neutralizing activity, (see examplesbelow) thus yielding information regarding which amino acids can bechanged and which must not be changed. In other words, based oninformation gathered from such routine experiments, one skilled in theart can readily determine the amino acid positions where furthersubstitutions should be avoided either alone or in combination withother mutations.

A number of scientific publications have been devoted to the predictionof secondary structure. See, Moult, 1996, Curr. Op. in Biotech.7:422-427; Chou et al., 1974, Biochem. 13:222-245; Chou et al., 1974,Biochemistry 113:211-222; Chou et al., 1978, Adv. Enzymol. Relat. AreasMol. Biol. 47:45-148; Chou et al., 1979, Ann. Rev. Biochem. 47:251-276;and Chou et al., 1979, Biophys. J. 26:367-384. Moreover, computerprograms are currently available to assist with predicting secondarystructure. One method of predicting secondary structure is based uponhomology modeling. For example, two polypeptides or proteins that have asequence identity of greater than 30%, or similarity greater than 40%can have similar structural topologies. The recent growth of the proteinstructural database (PDB) has provided enhanced predictability ofsecondary structure, including the potential number of folds within apolypeptide's or protein's structure. See, Holm et al., 1999, Nucl.Acid. Res. 27:244-247. It has been suggested (Brenner et al., 1997,Curr. Op. Struct. Biol. 7:369-376) that there are a limited number offolds in a given polypeptide or protein and that once a critical numberof structures have been resolved, structural prediction will becomedramatically more accurate.

Additional methods of predicting secondary structure include “threading”(Jones, 1997, Curr. Opin. Struct. Biol. 7:377-387; Sippl et al., 1996,Structure 4:15-19), “profile analysis” (Bowie et al., 1991, Science253:164-170; Gribskov et al., 1990, Meth. Enzym. 183:146-159; Gribskovet al., 1987, Proc. Nat. Acad. Sci. 84:4355-4358), and “evolutionarylinkage” (See, Holm, 1999, supra; and Brenner, 1997, supra).

In some embodiments, amino acid substitutions are made that: (1) reducesusceptibility to proteolysis, (2) reduce susceptibility to oxidation,(3) alter binding affinity for forming protein complexes, (4) alterligand or antigen binding affinities, and/or (4) confer or modify otherphysicochemical or functional properties on such polypeptides. Forexample, single or multiple amino acid substitutions (in certainembodiments, conservative amino acid substitutions) may be made in thenaturally-occurring sequence. Substitutions can be made in that portionof the antibody that lies outside the domain(s) forming intermolecularcontacts). In such embodiments, conservative amino acid substitutionscan be used that do not substantially change the structuralcharacteristics of the parent sequence (e.g., one or more replacementamino acids that do not disrupt the secondary structure thatcharacterizes the parent or native antibody). Examples of art-recognizedpolypeptide secondary and tertiary structures are described in Proteins,Structures and Molecular Principles (Creighton, Ed.), 1984, W. H. NewYork: Freeman and Company; Introduction to Protein Structure (Brandenand Tooze, eds.), 1991, New York: Garland Publishing; and Thornton etal., 1991, Nature 354:105, which are each incorporated herein byreference.

Additional preferred antibody variants include cysteine variants whereinone or more cysteine residues in the parent or native amino acidsequence are deleted from or substituted with another amino acid (e.g.,serine). Cysteine variants are useful, inter alia when antibodies mustbe refolded into a biologically active conformation. Cysteine variantsmay have fewer cysteine residues than the native antibody, and typicallyhave an even number to minimize interactions resulting from unpairedcysteines.

The heavy and light chains, variable regions domains and CDRs that aredisclosed can be used to prepare polypeptides that contain an antigenbinding region that can specifically bind to PAC1. For example, one ormore of the CDRs listed in Tables 5A and 5B can be incorporated into amolecule (e.g., a polypeptide) covalently or noncovalently to make animmunoadhesion. An immunoadhesion may incorporate the CDR(s) as part ofa larger polypeptide chain, may covalently link the CDR(s) to anotherpolypeptide chain, or may incorporate the CDR(s) noncovalently. TheCDR(s) enable the immunoadhesion to bind specifically to a particularantigen of interest (e.g., PAC1 or epitope thereof).

Mimetics (e.g., “peptide mimetics” or “peptidomimetics”) based upon thevariable region domains and CDRs that are described herein are alsoprovided. These analogs can be peptides, non-peptides or combinations ofpeptide and non-peptide regions. Fauchere, 1986, Adv. Drug Res. 15:29;Veber and Freidinger, 1985, TINS p. 392; and Evans et al., 1987, J. Med.Chem. 30:1229, which are incorporated herein by reference for anypurpose. Peptide mimetics that are structurally similar totherapeutically useful peptides may be used to produce a similartherapeutic or prophylactic effect. Such compounds are often developedwith the aid of computerized molecular modeling. Generally,peptidomimetics are proteins that are structurally similar to anantibody displaying a desired biological activity, such as here theability to specifically bind PAC1, but have one or more peptide linkagesoptionally replaced by a linkage selected from: —CH₂NH—, —CH₂S—,—CH₂—CH₂—, —CH—CH— (cis and trans), —COCH₂—, —CH(OH)CH₂—, and —CH₂SO—,by methods well known in the art. Systematic substitution of one or moreamino acids of a consensus sequence with a D-amino acid of the same type(e.g., D-lysine in place of L-lysine) may be used in certain embodimentsto generate more stable proteins. In addition, constrained peptidescomprising a consensus sequence or a substantially identical consensussequence variation may be generated by methods known in the art (Rizoand Gierasch, 1992, Ann. Rev. Biochem. 61:387), incorporated herein byreference), for example, by adding internal cysteine residues capable offorming intramolecular disulfide bridges which cyclize the peptide.

Derivatives of the antibodies that are described herein are alsoprovided. The derivatized antibodies can comprise any molecule orsubstance that imparts a desired property to the antibody or fragment,such as increased half-life in a particular use. The derivatizedantibody can comprise, for example, a detectable (or labeling) moiety(e.g., a radioactive, colorimetric, antigenic or enzymatic molecule, adetectable bead (such as a magnetic or electrodense (e.g., gold) bead),or a molecule that binds to another molecule (e.g., biotin orstreptavidin)), a therapeutic or diagnostic moiety (e.g., a radioactive,cytotoxic, or pharmaceutically active moiety), or a molecule thatincreases the suitability of the antibody for a particular use (e.g.,administration to a subject, such as a human subject, or other in vivoor in vitro uses). Examples of molecules that can be used to derivatizean antibody include albumin (e.g., human serum albumin) and polyethyleneglycol (PEG). Albumin-linked and PEGylated derivatives of antibodies canbe prepared using techniques well known in the art. Certain antibodiesinclude a pegylated single chain polypeptide as described herein. In oneembodiment, the antibody is conjugated or otherwise linked totransthyretin (TTR) or a TTR variant. The TTR or TTR variant can bechemically modified with, for example, a chemical selected from thegroup consisting of dextran, poly(n-vinyl pyrrolidone), polyethyleneglycols, propropylene glycol homopolymers, polypropylene oxide/ethyleneoxide co-polymers, polyoxyethylated polyols and polyvinyl alcohols.

Other derivatives include covalent or aggregative conjugates of PAC1binding proteins with other proteins or polypeptides, such as byexpression of recombinant fusion proteins comprising heterologouspolypeptides fused to the N-terminus or C-terminus of a PAC1 bindingprotein. For example, the conjugated peptide may be a heterologoussignal (or leader) polypeptide, e.g., the yeast alpha-factor leader, ora peptide such as an epitope tag. PAC1 antibody-containing fusionproteins can comprise peptides added to facilitate purification oridentification of the PAC1 binding protein (e.g., poly-His). A PAC1binding protein also can be linked to the FLAG peptide as described inHopp et al., 1988, Bio/Technology 6:1204; and U.S. Pat. No. 5,011,912.The FLAG peptide is highly antigenic and provides an epitope reversiblybound by a specific monoclonal antibody (mAb), enabling rapid assay andfacile purification of expressed recombinant protein. Reagents usefulfor preparing fusion proteins in which the FLAG peptide is fused to agiven polypeptide are commercially available (Sigma, St. Louis, Mo.).

Oligomers that contain one or more PAC1 binding proteins may be employedas PAC1 antagonists. Oligomers may be in the form of covalently-linkedor non-covalently-linked dimers, trimers, or higher oligomers. Oligomerscomprising two or more PAC1 binding proteins are contemplated for use,with one example being a homodimer. Other oligomers includeheterodimers, homotrimers, heterotrimers, homotetramers,heterotetramers, etc.

One embodiment is directed to oligomers comprising multiple PAC1-bindingpolypeptides joined via covalent or non-covalent interactions betweenpeptide moieties fused to the PAC1 binding proteins. Such peptides maybe peptide linkers (spacers), or peptides that have the property ofpromoting oligomerization. Leucine zippers and certain polypeptidesderived from antibodies are among the peptides that can promoteoligomerization of PAC1 binding proteins attached thereto, as describedin more detail below.

In particular embodiments, the oligomers comprise from two to four PAC1binding proteins. The PAC1 binding protein moieties of the oligomer maybe in any of the forms described above, e.g., variants or fragments.Preferably, the oligomers comprise PAC1 binding proteins that have PAC1binding activity.

In one embodiment, an oligomer is prepared using polypeptides derivedfrom immunoglobulins. Preparation of fusion proteins comprising certainheterologous polypeptides fused to various portions of antibody-derivedpolypeptides (including the Fc domain) has been described, e.g., byAshkenazi et al., 1991, Proc. Natl. Acad. Sci. USA 88:10535; Byrn etal., 1990, Nature 344:677; and Hollenbaugh et al., 1992 “Construction ofImmunoglobulin Fusion Proteins”, in Current Protocols in Immunology,Suppl. 4, pages 10.19.1-10.19.11.

One embodiment is directed to a dimer comprising two fusion proteinscreated by fusing a PAC1 binding protein to the Fc region of anantibody. The dimer can be made by, for example, inserting a gene fusionencoding the fusion protein into an appropriate expression vector,expressing the gene fusion in host cells transformed with therecombinant expression vector, and allowing the expressed fusion proteinto assemble much like antibody molecules, whereupon interchain disulfidebonds form between the Fc moieties to yield the dimer.

The term “Fc polypeptide” as used herein includes native and muteinforms of polypeptides derived from the Fc region of an antibody.Truncated forms of such polypeptides containing the hinge region thatpromotes dimerization also are included. Fusion proteins comprising Fcmoieties (and oligomers formed therefrom) offer the advantage of facilepurification by affinity chromatography over Protein A or Protein Gcolumns.

One suitable Fc polypeptide, described in PCT application WO 93/10151and U.S. Pat. Nos. 5,426,048 and 5,262,522, is a single chainpolypeptide extending from the N-terminal hinge region to the nativeC-terminus of the Fc region of a human IgG1 antibody. Another useful Fcpolypeptide is the Fc mutein described in U.S. Pat. No. 5,457,035, andin Baum et al., 1994, EMBO J. 13:3992-4001. The amino acid sequence ofthis mutein is identical to that of the native Fc sequence presented inWO 93/10151, except that amino acid 19 has been changed from Leu to Ala,amino acid 20 has been changed from Leu to Glu, and amino acid 22 hasbeen changed from Gly to Ala. The mutein exhibits reduced affinity forFc receptors.

In other embodiments, the variable portion of the heavy and/or lightchains of a PAC1 binding protein such as disclosed herein may besubstituted for the variable portion of an antibody heavy and/or lightchain.

Alternatively, the oligomer is a fusion protein comprising multiple PAC1binding proteins, with or without peptide linkers (spacer peptides).Among the suitable peptide linkers are those described in U.S. Pat. Nos.4,751,180 and 4,935,233.

Another method for preparing oligomeric PAC1 binding protein derivativesinvolves use of a leucine zipper. Leucine zipper domains are peptidesthat promote oligomerization of the proteins in which they are found.Leucine zippers were originally identified in several DNA-bindingproteins (Landschulz et al., 1988, Science 240:1759), and have sincebeen found in a variety of different proteins. Among the known leucinezippers are naturally occurring peptides and derivatives thereof thatdimerize or trimerize. Examples of leucine zipper domains suitable forproducing soluble oligomeric proteins are described in PCT applicationWO 94/10308, and the leucine zipper derived from lung surfactant proteinD (SPD) described in Hoppe et al., 1994, FEBS Letters 344:191, herebyincorporated by reference. The use of a modified leucine zipper thatallows for stable trimerization of a heterologous protein fused theretois described in Fanslow et al., 1994, Semin. Immunol. 6:267-278. In oneapproach, recombinant fusion proteins comprising a PAC1 binding proteinfragment or derivative fused to a leucine zipper peptide are expressedin suitable host cells, and the soluble oligomeric PAC1 binding proteinfragments or derivatives that form are recovered from the culturesupernatant.

In certain embodiments, the antibody has a K_(D) (equilibrium bindingaffinity) of less than 1 pM, 10 pM, 100 pM, 1 nM, 2 nM, 5 nM, 10 nM, 25nM or 50 nM.

Another aspect provides an antibody having a half-life of at least oneday in vitro or in vivo (e.g., when administered to a human subject). Inone embodiment, the antibody has a half-life of at least three days. Inanother embodiment, the antibody or portion thereof has a half-life offour days or longer. In another embodiment, the antibody or portionthereof has a half-life of eight days or longer. In another embodiment,the antibody or antigen-binding portion thereof is derivatized ormodified such that it has a longer half-life as compared to theunderivatized or unmodified antibody. In another embodiment, theantibody contains point mutations to increase serum half life, such asdescribed in WO 00/09560, published Feb. 24, 2000, incorporated byreference.

Glycosylation

The antibody may have a glycosylation pattern that is different oraltered from that found in the native species. As is known in the art,glycosylation patterns can depend on both the sequence of the protein(e.g., the presence or absence of particular glycosylation amino acidresidues, discussed below), or the host cell or organism in which theprotein is produced. Particular expression systems are discussed below.

Glycosylation of polypeptides is typically either N-linked or O-linked.N-linked refers to the attachment of the carbohydrate moiety to the sidechain of an asparagine residue. The tri-peptide sequencesasparagine-X-serine and asparagine-X-threonine, where X is any aminoacid except proline, are the recognition sequences for enzymaticattachment of the carbohydrate moiety to the asparagine side chain.Thus, the presence of either of these tri-peptide sequences in apolypeptide creates a potential glycosylation site. O-linkedglycosylation refers to the attachment of one of the sugarsN-acetylgalactosamine, galactose, or xylose, to a hydroxyamino acid,most commonly serine or threonine, although 5-hydroxyproline or5-hydroxylysine may also be used.

Addition of glycosylation sites to the antibody is convenientlyaccomplished by altering the amino acid sequence such that it containsone or more of the above-described tri-peptide sequences (for N-linkedglycosylation sites). The alteration may also be made by the additionof, or substitution by, one or more serine or threonine residues to thestarting sequence (for O-linked glycosylation sites). For ease, theantibody amino acid sequence may be altered through changes at the DNAlevel, particularly by mutating the DNA encoding the target polypeptideat preselected bases such that codons are generated that will translateinto the desired amino acids.

Another means of increasing the number of carbohydrate moieties on theantibody is by chemical or enzymatic coupling of glycosides to theprotein. These procedures are advantageous in that they do not requireproduction of the protein in a host cell that has glycosylationcapabilities for N- and O-linked glycosylation. Depending on thecoupling mode used, the sugar(s) may be attached to (a) arginine andhistidine, (b) free carboxyl groups, (c) free sulfhydryl groups such asthose of cysteine, (d) free hydroxyl groups such as those of serine,threonine, or hydroxyproline, (e) aromatic residues such as those ofphenylalanine, tyrosine, or tryptophan, or (f) the amide group ofglutamine. These methods are described in WO 87/05330 published Sep. 11,1987, and in Aplin and Wriston, 1981, CRC Crit. Rev, Biochem., pp.259-306.

Removal of carbohydrate moieties present on the starting antibody may beaccomplished chemically or enzymatically. Chemical deglycosylationrequires exposure of the protein to the compoundtrifluoromethanesulfonic acid, or an equivalent compound. This treatmentresults in the cleavage of most or all sugars except the linking sugar(N-acetylglucosamine or N-acetylgalactosamine), while leaving thepolypeptide intact. Chemical deglycosylation is described by Hakimuddinet al., 1987, Arch. Biochem. Biophys. 259:52 and by Edge et al., 1981,Anal. Biochem. 118:131. Enzymatic cleavage of carbohydrate moieties onpolypeptides can be achieved by the use of a variety of endo- andexo-glycosidases as described by Thotakura et al., 1987, Meth. Enzymol.138:350. Glycosylation at potential glycosylation sites may be preventedby the use of the compound tunicamycin as described by Duskin et al.,1982, J. Biol. Chem. 257:3105. Tunicamycin blocks the formation ofprotein-N-glycoside linkages.

Hence, aspects include glycosylation variants of the antibodies whereinthe number and/or type of glycosylation site(s) has been alteredcompared to the amino acid sequences of the parent polypeptide. Incertain embodiments, antibody protein variants comprise a greater or alesser number of N-linked glycosylation sites than the native antibody.An N-linked glycosylation site is characterized by the sequence:Asn-X-Ser or Asn-X-Thr, wherein the amino acid residue designated as Xmay be any amino acid residue except proline. The substitution of aminoacid residues to create this sequence provides a potential new site forthe addition of an N-linked carbohydrate chain. Alternatively,substitutions that eliminate or alter this sequence will preventaddition of an N-linked carbohydrate chain present in the nativepolypeptide. For example, the glycosylation can be reduced by thedeletion of an Asn or by substituting the Asn with a different aminoacid. In other embodiments, one or more new N-linked sites are created.Antibodies typically have a N-linked glycosylation site in the Fcregion.

Labels and Effector Groups

In some embodiments, the antigen-binding comprises one or more labels.The term “labeling group” or “label” means any detectable label.Examples of suitable labeling groups include, but are not limited to,the following: radioisotopes or radionuclides (e.g., ³H, ¹⁴C, ¹⁵N, ³⁵S,⁹⁰Y, ⁹⁹Tc, ¹¹¹In, ¹²⁵I, ¹³¹I), fluorescent groups (e.g., FITC,rhodamine, lanthanide phosphors), enzymatic groups (e.g., horseradishperoxidase, β-galactosidase, luciferase, alkaline phosphatase),chemiluminescent groups, biotinyl groups, or predetermined polypeptideepitopes recognized by a secondary reporter (e.g., leucine zipper pairsequences, binding sites for secondary antibodies, metal bindingdomains, epitope tags). In some embodiments, the labeling group iscoupled to the antibody via spacer arms of various lengths to reducepotential steric hindrance. Various methods for labeling proteins areknown in the art and may be used as is seen fit.

The term “effector group” means any group coupled to an antibody thatacts as a cytotoxic agent. Examples for suitable effector groups areradioisotopes or radionuclides (e.g., ³H, ¹⁴C, ¹⁵N, ³⁵S, ⁹⁰Y, ⁹⁹Tc,¹¹¹In, ¹²⁵I, ¹³¹I). Other suitable groups include toxins, therapeuticgroups, or chemotherapeutic groups. Examples of suitable groups includecalicheamicin, auristatins, geldanamycin and maytansine. In someembodiments, the effector group is coupled to the antibody via spacerarms of various lengths to reduce potential steric hindrance.

In general, labels fall into a variety of classes, depending on theassay in which they are to be detected: a) isotopic labels, which may beradioactive or heavy isotopes; b) magnetic labels (e.g., magneticparticles); c) redox active moieties; d) optical dyes; enzymatic groups(e.g. horseradish peroxidase, β-galactosidase, luciferase, alkalinephosphatase); e) biotinylated groups; and f) predetermined polypeptideepitopes recognized by a secondary reporter (e.g., leucine zipper pairsequences, binding sites for secondary antibodies, metal bindingdomains, epitope tags, etc.). In some embodiments, the labeling group iscoupled to the antibody via spacer arms of various lengths to reducepotential steric hindrance. Various methods for labeling proteins areknown in the art.

Specific labels include optical dyes, including, but not limited to,chromophores, phosphors and fluorophores, with the latter being specificin many instances. Fluorophores can be either “small molecule” fluores,or proteinaceous fluores.

By “fluorescent label” is meant any molecule that may be detected viaits inherent fluorescent properties. Suitable fluorescent labelsinclude, but are not limited to, fluorescein, rhodamine,tetramethylrhodamine, eosin, erythrosin, coumarin, methyl-coumarins,pyrene, Malacite green, stilbene, Lucifer Yellow, Cascade BlueJ, TexasRed, IAEDANS, EDANS, BODIPY FL, LC Red 640, Cy 5, Cy 5.5, LC Red 705,Oregon green, the Alexa-Fluor dyes (Alexa Fluor 350, Alexa Fluor 430,Alexa Fluor 488, Alexa Fluor 546, Alexa Fluor 568, Alexa Fluor 594,Alexa Fluor 633, Alexa Fluor 647, Alexa Fluor 660, Alexa Fluor 680),Cascade Blue, Cascade Yellow and R-phycoerythrin (PE) (Molecular Probes,Eugene, Oreg.), FITC, Rhodamine, and Texas Red (Pierce, Rockford, Ill.),Cy5, Cy5.5, Cy7 (Amersham Life Science, Pittsburgh, Pa.). Suitableoptical dyes, including fluorophores, are described in MOLECULAR PROBESHANDBOOK by Richard P. Haugland, hereby expressly incorporated byreference.

Suitable proteinaceous fluorescent labels also include, but are notlimited to, green fluorescent protein, including a Renilla, Ptilosarcus,or Aequorea species of GFP (Chalfie et al., 1994, Science 263:802-805),EGFP (Clontech Labs., Inc., Genbank Accession Number U55762), bluefluorescent protein (BFP, Quantum Biotechnologies, Inc., Quebec, Canada;Stauber, 1998, Biotechniques 24:462-471; Heim et al., 1996, Curr. Biol.6:178-182), enhanced yellow fluorescent protein (EYFP, Clontech Labs.,Inc.), luciferase (Ichiki et al., 1993, J. Immunol. 150:5408-5417), βgalactosidase (Nolan et al., 1988, Proc. Natl. Acad. Sci. U.S.A.85:2603-2607) and Renilla (WO92/15673, WO95/07463, WO98/14605,WO98/26277, WO99/49019, U.S. Pat. Nos. 5,292,658, 5,418,155, 5,683,888,5,741,668, 5,777,079, 5,804,387, 5,874,304, 5,876,995, 5,925,558).

Nucleic Acids

An aspect further provides nucleic acids that hybridize to other nucleicacids (e.g., nucleic acids comprising a nucleotide sequence listed inTables 1A, 1B, 3A and 3B) under particular hybridization conditions.Methods for hybridizing nucleic acids are well-known in the art. See,e.g., Current Protocols in Molecular Biology, John Wiley & Sons, N.Y.(1989), 6.3.1-6.3.6. As defined herein, a moderately stringenthybridization condition uses a prewashing solution containing 5× sodiumchloride/sodium citrate (SSC), 0.5% SDS, 1.0 mM EDTA (pH 8.0),hybridization buffer of about 50% formamide, 6×SSC, and a hybridizationtemperature of 55° C. (or other similar hybridization solutions, such asone containing about 50% formamide, with a hybridization temperature of42° C.), and washing conditions of 60° C., in 0.5×SSC, 0.1% SDS. Astringent hybridization condition hybridizes in 6×SSC at 45° C.,followed by one or more washes in 0.1×SSC, 0.2% SDS at 68° C.Furthermore, one of skill in the art can manipulate the hybridizationand/or washing conditions to increase or decrease the stringency ofhybridization such that nucleic acids comprising nucleotide sequencesthat are at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99%identical to each other typically remain hybridized to each other.

The basic parameters affecting the choice of hybridization conditionsand guidance for devising suitable conditions are set forth by, forexample, Sambrook, Fritsch, and Maniatis (2001, Molecular Cloning: ALaboratory Manual, Cold Spring Harbor Laboratory Press, Cold SpringHarbor, N.Y., supra; and Current Protocols in Molecular Biology, 1995,Ausubel et al., eds., John Wiley & Sons, Inc., sections 2.10 and6.3-6.4), and can be readily determined by those having ordinary skillin the art based on, e.g., the length and/or base composition of thenucleic acid.

Changes can be introduced by mutation into a nucleic acid, therebyleading to changes in the amino acid sequence of a polypeptide (e.g., anantibody or antibody derivative) that it encodes. Mutations can beintroduced using any technique known in the art. In one embodiment, oneor more particular amino acid residues are changed using, for example, asite-directed mutagenesis protocol. In another embodiment, one or morerandomly selected residues is changed using, for example, a randommutagenesis protocol. However it is made, a mutant polypeptide can beexpressed and screened for a desired property.

Mutations can be introduced into a nucleic acid without significantlyaltering the biological activity of a polypeptide that it encodes. Forexample, one can make nucleotide substitutions leading to amino acidsubstitutions at non-essential amino acid residues. Alternatively, oneor more mutations can be introduced into a nucleic acid that selectivelychanges the biological activity of a polypeptide that it encodes. Forexample, the mutation can quantitatively or qualitatively change thebiological activity. Examples of quantitative changes includeincreasing, reducing or eliminating the activity. Examples ofqualitative changes include changing the antigen specificity of anantibody. In one embodiment, a nucleic acid encoding any antibodydescribed herein can be mutated to alter the amino acid sequence usingmolecular biology techniques that are well-established in the art.

Another aspect provides nucleic acid molecules that are suitable for useas primers or hybridization probes for the detection of nucleic acidsequences. A nucleic acid molecule can comprise only a portion of anucleic acid sequence encoding a full-length polypeptide, for example, afragment that can be used as a probe or primer or a fragment encoding anactive portion (e.g., a PAC1 binding portion) of a polypeptide.

Probes based on the sequence of a nucleic acid can be used to detect thenucleic acid or similar nucleic acids, for example, transcripts encodinga polypeptide. The probe can comprise a label group, e.g., aradioisotope, a fluorescent compound, an enzyme, or an enzyme co-factor.Such probes can be used to identify a cell that expresses thepolypeptide.

Another aspect provides vectors comprising a nucleic acid encoding apolypeptide or a portion thereof (e.g., a fragment containing one ormore CDRs or one or more variable region domains). Examples of vectorsinclude, but are not limited to, plasmids, viral vectors, non-episomalmammalian vectors and expression vectors, for example, recombinantexpression vectors. The recombinant expression vectors can comprise anucleic acid in a form suitable for expression of the nucleic acid in ahost cell. The recombinant expression vectors include one or moreregulatory sequences, selected on the basis of the host cells to be usedfor expression, which is operably linked to the nucleic acid sequence tobe expressed. Regulatory sequences include those that directconstitutive expression of a nucleotide sequence in many types of hostcells (e.g., SV40 early gene enhancer, Rous sarcoma virus promoter andcytomegalovirus promoter), those that direct expression of thenucleotide sequence only in certain host cells (e.g., tissue-specificregulatory sequences, see, Voss et al., 1986, Trends Biochem. Sci.11:287, Maniatis et al., 1987, Science 236:1237, incorporated byreference herein in their entireties), and those that direct inducibleexpression of a nucleotide sequence in response to particular treatmentor condition (e.g., the metallothionin promoter in mammalian cells andthe tet-responsive and/or streptomycin responsive promoter in bothprokaryotic and eukaryotic systems (see, id.). It will be appreciated bythose skilled in the art that the design of the expression vector candepend on such factors as the choice of the host cell to be transformed,the level of expression of protein desired, etc. The expression vectorscan be introduced into host cells to thereby produce proteins orpeptides, including fusion proteins or peptides, encoded by nucleicacids as described herein.

Another aspect provides host cells into which a recombinant expressionvector has been introduced. A host cell can be any prokaryotic cell (forexample, E. coli) or eukaryotic cell (for example, yeast, insect, ormammalian cells (e.g., CHO cells)). Vector DNA can be introduced intoprokaryotic or eukaryotic cells via conventional transformation ortransfection techniques. For stable transfection of mammalian cells, itis known that, depending upon the expression vector and transfectiontechnique used, only a small fraction of cells may integrate the foreignDNA into their genome. In order to identify and select these integrants,a gene that encodes a selectable marker (e.g., for resistance toantibiotics) is generally introduced into the host cells along with thegene of interest. Preferred selectable markers include those whichconfer resistance to drugs, such as G418, hygromycin and methotrexate.Cells stably transfected with the introduced nucleic acid can beidentified by drug selection (e.g., cells that have incorporated theselectable marker gene will survive, while the other cells die), amongother methods

Preparing Antibodies

Non-human antibodies that are provided can be, for example, derived fromany antibody-producing animal, such as mouse, rat, rabbit, goat, donkey,or non-human primate (such as monkey (e.g., cynomolgus or rhesus monkey)or ape (e.g., chimpanzee)). Non-human antibodies can be used, forinstance, in vitro cell culture and cell-culture based applications, orany other application where an immune response to the antibody does notoccur or is insignificant, can be prevented, is not a concern, or isdesired. In certain embodiments, the antibodies may be produced byimmunizing animals using methods known in the art, as described above.The antibodies may be polyclonal, monoclonal, or may be synthesized inhost cells by expressing recombinant DNA. Fully human antibodies may beprepared as described above by immunizing transgenic animals containinghuman immunoglobulin loci or by selecting a phage display library thatis expressing a repertoire of human antibodies.

The monoclonal antibodies (mAbs) can be produced by a variety oftechniques, including conventional monoclonal antibody methodology,e.g., the standard somatic cell hybridization technique of Kohler andMilstein, 1975, Nature 256:495. Alternatively, other techniques forproducing monoclonal antibodies can be employed, for example, the viralor oncogenic transformation of B-lymphocytes. One suitable animal systemfor preparing hybridomas is the murine system, which is a very wellestablished procedure. Immunization protocols and techniques forisolation of immunized splenocytes for fusion are known in the art andillustrative approaches are described in the Examples, below. For suchprocedures, B cells from immunized mice are typically fused with asuitable immortalized fusion partner, such as a murine myeloma cellline. If desired, rats or other mammals besides can be immunized insteadof mice and B cells from such animals can be fused with the murinemyeloma cell line to form hybridomas. Alternatively, a myeloma cell linefrom a source other than mouse may be used. Fusion procedures for makinghybridomas also are well known.

The single chain antibodies that are provided may be formed by linkingheavy and light chain variable domain (Fv region) fragments via an aminoacid bridge (short peptide linker), resulting in a single polypeptidechain. Such single-chain Fvs (scFvs) may be prepared by fusing DNAencoding a peptide linker between DNAs encoding the two variable domainpolypeptides (V_(L) and V_(H)). The resulting polypeptides can fold backon themselves to form antigen-binding monomers, or they can formmultimers (e.g., dimers, trimers, or tetramers), depending on the lengthof a flexible linker between the two variable domains (Kortt et al.,1997, Prot. Eng. 10:423; Kortt et al., 2001, Biomol. Eng. 18:95-108). Bycombining different V_(L) and V_(H)-comprising polypeptides, one canform multimeric scFvs that bind to different epitopes (Kriangkum et al.,2001, Biomol. Eng. 18:31-40). Techniques developed for the production ofsingle chain antibodies include those described in U.S. Pat. No.4,946,778; Bird, 1988, Science 242:423; Huston et al., 1988, Proc. Natl.Acad. Sci. U.S.A. 85:5879; Ward et al., 1989, Nature 334:544, de Graafet al., 2002, Methods Mol Biol. 178:379-387. Single chain antibodiesderived from antibodies provided herein include, but are not limited toscFvs comprising the variable domain combinations of the heavy and lightchain variable regions depicted in Tables 4A and 4B, or combinations oflight and heavy chain variable domains which include CDRs depicted inTables 5A and 5B.

Antibodies provided herein that are of one subclass can be changed toantibodies from a different subclass using subclass switching methods.Thus, IgG antibodies may be derived from an IgM antibody, for example,and vice versa. Such techniques allow the preparation of new antibodiesthat possess the antigen binding properties of a given antibody (theparent antibody), but also exhibit biological properties associated withan antibody isotype or subclass different from that of the parentantibody. Recombinant DNA techniques may be employed. Cloned DNAencoding particular antibody polypeptides may be employed in suchprocedures, e.g., DNA encoding the constant domain of an antibody of thedesired isotype. See, e.g., Lantto et al., 2002, Methods Mol. Biol.178:303-316.

Accordingly, the antibodies that are provided include those comprising,for example, the variable domain combinations described, supra., havinga desired isotype (for example, IgA, IgG1, IgG2, IgG3, IgG4, IgE, andIgD) as well as Fab or F(ab′)₂ fragments thereof. Moreover, if an IgG4is desired, it may also be desired to introduce a point mutation(CPSCP->CPPCP) (SEQ ID NOS 193-194, respectively, in order ofappearance) in the hinge region as described in Bloom et al., 1997,Protein Science 6:407, incorporated by reference herein) to alleviate atendency to form intra-H chain disulfide bonds that can lead toheterogeneity in the IgG4 antibodies.

Moreover, techniques for deriving antibodies having different properties(i.e., varying affinities for the antigen to which they bind) are alsoknown. One such technique, referred to as chain shuffling, involvesdisplaying immunoglobulin variable domain gene repertoires on thesurface of filamentous bacteriophage, often referred to as phagedisplay. Chain shuffling has been used to prepare high affinityantibodies to the hapten 2-phenyloxazol-5-one, as described by Marks etal., 1992, BioTechnology 10:779.

Conservative modifications may be made to the heavy and light chainvariable regions described in Tables 4A and 4B, or the CDRs described inTables 5A and 5B (and corresponding modifications to the encodingnucleic acids) to produce a PAC1 binding protein having certaindesirable functional and biochemical characteristics. Methods forachieving such modifications are described above.

PAC1 antibodies may be further modified in various ways. For example, ifthey are to be used for therapeutic purposes, they may be conjugatedwith polyethylene glycol (pegylated) to prolong the serum half-life orto enhance protein delivery. Alternatively, the V region of the subjectantibodies or fragments thereof may be fused with the Fc region of adifferent antibody molecule. The Fc region used for this purpose may bemodified so that it does not bind complement, thus reducing thelikelihood of inducing cell lysis in the patient when the fusion proteinis used as a therapeutic agent. In addition, the subject antibodies orfunctional fragments thereof may be conjugated with human serum albuminto enhance the serum half-life of the antibody or antigen bindingfragment thereof. Another useful fusion partner for the antibodies orfragments thereof is transthyretin (TTR). TTR has the capacity to form atetramer, thus an antibody-TTR fusion protein can form a multivalentantibody which may increase its binding avidity.

Alternatively, substantial modifications in the functional and/orbiochemical characteristics of the antibodies described herein may beachieved by creating substitutions in the amino acid sequence of theheavy and light chains that differ significantly in their effect onmaintaining (a) the structure of the molecular backbone in the area ofthe substitution, for example, as a sheet or helical conformation, (b)the charge or hydrophobicity of the molecule at the target site, or (c)the bulkiness of the side chain. A “conservative amino acidsubstitution” may involve a substitution of a native amino acid residuewith a nonnative residue that has little or no effect on the polarity orcharge of the amino acid residue at that position. See, Table 7, supra.Furthermore, any native residue in the polypeptide may also besubstituted with alanine, as has been previously described for alaninescanning mutagenesis.

Amino acid substitutions (whether conservative or non-conservative) ofthe subject antibodies can be implemented by those skilled in the art byapplying routine techniques. Amino acid substitutions can be used toidentify important residues of the antibodies provided herein, or toincrease or decrease the affinity of these antibodies for human PAC1 orfor modifying the binding affinity of other antibodies described herein.

Methods of Expressing Antibodies

Expression systems and constructs in the form of plasmids, expressionvectors, transcription or expression cassettes that comprise at leastone polynucleotide as described above are also provided herein, as wellhost cells comprising such expression systems or constructs.

The antibodies provided herein may be prepared by any of a number ofconventional techniques. For example, PAC1 antibodies may be produced byrecombinant expression systems, using any technique known in the art.See, e.g., Monoclonal Antibodies, Hybridomas: A New Dimension inBiological Analyses, Kennet et al. (eds.) Plenum Press, New York (1980);and Antibodies: A Laboratory Manual, Harlow and Lane (eds.), Cold SpringHarbor Laboratory Press, Cold Spring Harbor, N.Y. (1988).

Antibodies can be expressed in hybridoma cell lines (e.g., in particularantibodies may be expressed in hybridomas) or in cell lines other thanhybridomas. Expression constructs encoding the antibodies can be used totransform a mammalian, insect or microbial host cell. Transformation canbe performed using any known method for introducing polynucleotides intoa host cell, including, for example packaging the polynucleotide in avirus or bacteriophage and transducing a host cell with the construct bytransfection procedures known in the art, as exemplified by U.S. Pat.Nos. 4,399,216; 4,912,040; 4,740,461; 4,959,455. The optimaltransformation procedure used will depend upon which type of host cellis being transformed. Methods for introduction of heterologouspolynucleotides into mammalian cells are well known in the art andinclude, but are not limited to, dextran-mediated transfection, calciumphosphate precipitation, polybrene mediated transfection, protoplastfusion, electroporation, encapsulation of the polynucleotide(s) inliposomes, mixing nucleic acid with positively-charged lipids, anddirect microinjection of the DNA into nuclei.

Recombinant expression constructs typically comprise a nucleic acidmolecule encoding a polypeptide comprising one or more of the following:one or more CDRs provided herein; a light chain constant region; a lightchain variable region; a heavy chain constant region (e.g., C_(H)1,C_(H)2 and/or C_(H)3); and/or another scaffold portion of a PAC1antibody. These nucleic acid sequences are inserted into an appropriateexpression vector using standard ligation techniques. In one embodiment,the heavy or light chain constant region is appended to the C-terminusof the anti-PAC1-specific heavy or light chain variable region and isligated into an expression vector. The vector is typically selected tobe functional in the particular host cell employed (i.e., the vector iscompatible with the host cell machinery, permitting amplification and/orexpression of the gene can occur). In some embodiments, vectors are usedthat employ protein-fragment complementation assays using proteinreporters, such as dihydrofolate reductase (see, for example, U.S. Pat.No. 6,270,964, which is hereby incorporated by reference). Suitableexpression vectors can be purchased, for example, from Invitrogen LifeTechnologies or BD Biosciences (formerly “Clontech”). Other usefulvectors for cloning and expressing the antibodies and fragments includethose described in Bianchi and McGrew, 2003, Biotech. Biotechnol.Bioeng. 84:439-44, which is hereby incorporated by reference. Additionalsuitable expression vectors are discussed, for example, in MethodsEnzymol., vol. 185 (D. V. Goeddel, ed.), 1990, New York: Academic Press.

Typically, expression vectors used in any of the host cells will containsequences for plasmid maintenance and for cloning and expression ofexogenous nucleotide sequences. Such sequences, collectively referred toas “flanking sequences” in certain embodiments will typically includeone or more of the following nucleotide sequences: a promoter, one ormore enhancer sequences, an origin of replication, a transcriptionaltermination sequence, a complete intron sequence containing a donor andacceptor splice site, a sequence encoding a leader sequence forpolypeptide secretion, a ribosome binding site, a polyadenylationsequence, a polylinker region for inserting the nucleic acid encodingthe polypeptide to be expressed, and a selectable marker element. Eachof these sequences is discussed below.

Optionally, the vector may contain a “tag”-encoding sequence, i.e., anoligonucleotide molecule located at the 5′ or 3′ end of the PAC1 bindingprotein coding sequence; the oligonucleotide sequence encodes polyHis(such as hexaHis (SEQ ID NO: 195)), or another “tag” such as FLAG®, HA(hemaglutinin influenza virus), or myc, for which commercially availableantibodies exist. This tag is typically fused to the polypeptide uponexpression of the polypeptide, and can serve as a means for affinitypurification or detection of the PAC1 binding protein from the hostcell. Affinity purification can be accomplished, for example, by columnchromatography using antibodies against the tag as an affinity matrix.Optionally, the tag can subsequently be removed from the purified PAC1binding protein by various means such as using certain peptidases forcleavage.

Flanking sequences may be homologous (i.e., from the same species and/orstrain as the host cell), heterologous (i.e., from a species other thanthe host cell species or strain), hybrid (i.e., a combination offlanking sequences from more than one source), synthetic or native. Assuch, the source of a flanking sequence may be any prokaryotic oreukaryotic organism, any vertebrate or invertebrate organism, or anyplant, provided that the flanking sequence is functional in, and can beactivated by, the host cell machinery.

Flanking sequences useful in the vectors may be obtained by any ofseveral methods well known in the art. Typically, flanking sequencesuseful herein will have been previously identified by mapping and/or byrestriction endonuclease digestion and can thus be isolated from theproper tissue source using the appropriate restriction endonucleases. Insome cases, the full nucleotide sequence of a flanking sequence may beknown. Here, the flanking sequence may be synthesized using the methodsdescribed herein for nucleic acid synthesis or cloning.

Whether all or only a portion of the flanking sequence is known, it maybe obtained using polymerase chain reaction (PCR) and/or by screening agenomic library with a suitable probe such as an oligonucleotide and/orflanking sequence fragment from the same or another species. Where theflanking sequence is not known, a fragment of DNA containing a flankingsequence may be isolated from a larger piece of DNA that may contain,for example, a coding sequence or even another gene or genes. Isolationmay be accomplished by restriction endonuclease digestion to produce theproper DNA fragment followed by isolation using agarose gelpurification, Qiagen® column chromatography (Chatsworth, Calif.), orother methods known to the skilled artisan. The selection of suitableenzymes to accomplish this purpose will be readily apparent to one ofordinary skill in the art.

An origin of replication is typically a part of those prokaryoticexpression vectors purchased commercially, and the origin aids in theamplification of the vector in a host cell. If the vector of choice doesnot contain an origin of replication site, one may be chemicallysynthesized based on a known sequence, and ligated into the vector. Forexample, the origin of replication from the plasmid pBR322 (New EnglandBiolabs, Beverly, Mass.) is suitable for most gram-negative bacteria,and various viral origins (e.g., SV40, polyoma, adenovirus, vesicularstomatitus virus (VSV), or papillomaviruses such as HPV or BPV) areuseful for cloning vectors in mammalian cells. Generally, the origin ofreplication component is not needed for mammalian expression vectors(for example, the SV40 origin is often used only because it alsocontains the virus early promoter).

A transcription termination sequence is typically located 3′ to the endof a polypeptide coding region and serves to terminate transcription.Usually, a transcription termination sequence in prokaryotic cells is aG-C rich fragment followed by a poly-T sequence. While the sequence iseasily cloned from a library or even purchased commercially as part of avector, it can also be readily synthesized using methods for nucleicacid synthesis such as those described herein.

A selectable marker gene encodes a protein necessary for the survivaland growth of a host cell grown in a selective culture medium. Typicalselection marker genes encode proteins that (a) confer resistance toantibiotics or other toxins, e.g., ampicillin, tetracycline, orkanamycin for prokaryotic host cells; (b) complement auxotrophicdeficiencies of the cell; or (c) supply critical nutrients not availablefrom complex or defined media. Specific selectable markers are thekanamycin resistance gene, the ampicillin resistance gene, and thetetracycline resistance gene. Advantageously, a neomycin resistance genemay also be used for selection in both prokaryotic and eukaryotic hostcells.

Other selectable genes may be used to amplify the gene that will beexpressed. Amplification is the process wherein genes that are requiredfor production of a protein critical for growth or cell survival arereiterated in tandem within the chromosomes of successive generations ofrecombinant cells. Examples of suitable selectable markers for mammaliancells include dihydrofolate reductase (DHFR) and promoterless thymidinekinase genes. Mammalian cell transformants are placed under selectionpressure wherein only the transformants are uniquely adapted to surviveby virtue of the selectable gene present in the vector. Selectionpressure is imposed by culturing the transformed cells under conditionsin which the concentration of selection agent in the medium issuccessively increased, thereby leading to the amplification of both theselectable gene and the DNA that encodes another gene, such as anantibody that binds to PAC1. As a result, increased quantities of apolypeptide such as an antibody are synthesized from the amplified DNA.

A ribosome-binding site is usually necessary for translation initiationof mRNA and is characterized by a Shine-Dalgarno sequence (prokaryotes)or a Kozak sequence (eukaryotes). The element is typically located 3′ tothe promoter and 5′ to the coding sequence of the polypeptide to beexpressed.

In some cases, such as where glycosylation is desired in a eukaryotichost cell expression system, one may manipulate the various pre- orpro-sequences to improve glycosylation or yield. For example, one mayalter the peptidase cleavage site of a particular signal peptide, or addprosequences, which also may affect glycosylation. The final proteinproduct may have, in the −1 position (relative to the first amino acidof the mature protein), one or more additional amino acids incident toexpression, which may not have been totally removed. For example, thefinal protein product may have one or two amino acid residues found inthe peptidase cleavage site, attached to the amino-terminus.Alternatively, use of some enzyme cleavage sites may result in aslightly truncated form of the desired polypeptide, if the enzyme cutsat such area within the mature polypeptide.

Expression and cloning will typically contain a promoter that isrecognized by the host organism and operably linked to the moleculeencoding a PAC1 binding protein. Promoters are untranscribed sequenceslocated upstream (i.e., 5′) to the start codon of a structural gene(generally within about 100 to 1000 bp) that control transcription ofthe structural gene. Promoters are conventionally grouped into one oftwo classes: inducible promoters and constitutive promoters. Induciblepromoters initiate increased levels of transcription from DNA undertheir control in response to some change in culture conditions, such asthe presence or absence of a nutrient or a change in temperature.Constitutive promoters, on the other hand, uniformly transcribe a geneto which they are operably linked, that is, with little or no controlover gene expression. A large number of promoters, recognized by avariety of potential host cells, are well known. A suitable promoter isoperably linked to the DNA encoding heavy chain or light chaincomprising a PAC1 binding protein by removing the promoter from thesource DNA by restriction enzyme digestion and inserting the desiredpromoter sequence into the vector.

Suitable promoters for use with yeast hosts are also well known in theart. Yeast enhancers are advantageously used with yeast promoters.Suitable promoters for use with mammalian host cells are well known andinclude, but are not limited to, those obtained from the genomes ofviruses such as polyoma virus, fowlpox virus, adenovirus (such asAdenovirus 2), bovine papilloma virus, avian sarcoma virus,cytomegalovirus, retroviruses, hepatitis-B virus, and Simian Virus 40(SV40). Other suitable mammalian promoters include heterologousmammalian promoters, for example, heat-shock promoters and the actinpromoter.

Additional promoters which may be of interest include, but are notlimited to: SV40 early promoter (Benoist and Chambon, 1981, Nature290:304-310); CMV promoter (Thornsen et al., 1984, Proc. Natl. Acad.U.S.A. 81:659-663); the promoter contained in the 3′ long terminalrepeat of Rous sarcoma virus (Yamamoto et al., 1980, Cell 22:787-797);herpes thymidine kinase promoter (Wagner et al., 1981, Proc. Natl. Acad.Sci. U.S.A. 78:1444-1445); promoter and regulatory sequences from themetallothionine gene (Prinster et al., 1982, Nature 296:39-42); andprokaryotic promoters such as the beta-lactamase promoter(Villa-Kamaroff et al., 1978, Proc. Natl. Acad. Sci. U.S.A.75:3727-3731); or the tac promoter (DeBoer et al., 1983, Proc. Natl.Acad. Sci. U.S.A. 80:21-25). Also of interest are the following animaltranscriptional control regions, which exhibit tissue specificity andhave been utilized in transgenic animals: the elastase I gene controlregion that is active in pancreatic acinar cells (Swift et al., 1984,Cell 38:639-646; Ornitz et al., 1986, Cold Spring Harbor Symp. Quant.Biol. 50:399-409; MacDonald, 1987, Hepatology 7:425-515); the insulingene control region that is active in pancreatic beta cells (Hanahan,1985, Nature 315:115-122); the immunoglobulin gene control region thatis active in lymphoid cells (Grosschedl et al., 1984, Cell 38:647-658;Adames et al., 1985, Nature 318:533-538; Alexander et al., 1987, Mol.Cell. Biol. 7:1436-1444); the mouse mammary tumor virus control regionthat is active in testicular, breast, lymphoid and mast cells (Leder etal., 1986, Cell 45:485-495); the albumin gene control region that isactive in liver (Pinkert et al., 1987, Genes and Devel. 1:268-276); thealpha-feto-protein gene control region that is active in liver (Krumlaufet al., 1985, Mol. Cell. Biol. 5:1639-1648; Hammer et al., 1987, Science253:53-58); the alpha 1-antitrypsin gene control region that is activein liver (Kelsey et al., 1987, Genes and Devel. 1:161-171); thebeta-globin gene control region that is active in myeloid cells (Mogramet al., 1985, Nature 315:338-340; Kollias et al., 1986, Cell 46:89-94);the myelin basic protein gene control region that is active inoligodendrocyte cells in the brain (Readhead et al., 1987, Cell48:703-712); the myosin light chain-2 gene control region that is activein skeletal muscle (Sani, 1985, Nature 314:283-286); and thegonadotropic releasing hormone gene control region that is active in thehypothalamus (Mason et al., 1986, Science 234:1372-1378).

An enhancer sequence may be inserted into the vector to increasetranscription of DNA encoding light chain or heavy chain comprising ahuman PAC1 binding protein by higher eukaryotes. Enhancers arecis-acting elements of DNA, usually about 10-300 bp in length, that acton the promoter to increase transcription. Enhancers are relativelyorientation and position independent, having been found at positionsboth 5′ and 3′ to the transcription unit. Several enhancer sequencesavailable from mammalian genes are known (e.g., globin, elastase,albumin, alpha-feto-protein and insulin). Typically, however, anenhancer from a virus is used. The SV40 enhancer, the cytomegalovirusearly promoter enhancer, the polyoma enhancer, and adenovirus enhancersknown in the art are exemplary enhancing elements for the activation ofeukaryotic promoters. While an enhancer may be positioned in the vectoreither 5′ or 3′ to a coding sequence, it is typically located at a site5′ from the promoter. A sequence encoding an appropriate native orheterologous signal sequence (leader sequence or signal peptide) can beincorporated into an expression vector, to promote extracellularsecretion of the antibody. The choice of signal peptide or leaderdepends on the type of host cells in which the antibody is to beproduced, and a heterologous signal sequence can replace the nativesignal sequence. Examples of signal peptides that are functional inmammalian host cells include the following: the signal sequence forinterleukin-7 (IL-7) described in U.S. Pat. No. 4,965,195; the signalsequence for interleukin-2 receptor described in Cosman et al., 1984,Nature 312:768; the interleukin-4 receptor signal peptide described inEP Patent No. 0367 566; the type I interleukin-1 receptor signal peptidedescribed in U.S. Pat. No. 4,968,607; the type II interleukin-1 receptorsignal peptide described in EP Patent No. 0 460 846.

The expression vectors that are provided may be constructed from astarting vector such as a commercially available vector. Such vectorsmay or may not contain all of the desired flanking sequences. Where oneor more of the flanking sequences described herein are not alreadypresent in the vector, they may be individually obtained and ligatedinto the vector. Methods used for obtaining each of the flankingsequences are well known to one skilled in the art.

After the vector has been constructed and a nucleic acid moleculeencoding light chain, a heavy chain, or a light chain and a heavy chaincomprising a PAC1 antigen binding sequence has been inserted into theproper site of the vector, the completed vector may be inserted into asuitable host cell for amplification and/or polypeptide expression. Thetransformation of an expression vector for an antibody into a selectedhost cell may be accomplished by well known methods includingtransfection, infection, calcium phosphate co-precipitation,electroporation, microinjection, lipofection, DEAE-dextran mediatedtransfection, or other known techniques. The method selected will inpart be a function of the type of host cell to be used. These methodsand other suitable methods are well known to the skilled artisan, andare set forth, for example, in Sambrook et al., 2001, supra.

A host cell, when cultured under appropriate conditions, synthesizes anantibody that can subsequently be collected from the culture medium (ifthe host cell secretes it into the medium) or directly from the hostcell producing it (if it is not secreted). The selection of anappropriate host cell will depend upon various factors, such as desiredexpression levels, polypeptide modifications that are desirable ornecessary for activity (such as glycosylation or phosphorylation) andease of folding into a biologically active molecule.

Mammalian cell lines available as hosts for expression are well known inthe art and include, but are not limited to, immortalized cell linesavailable from the American Type Culture Collection (ATCC), includingbut not limited to Chinese hamster ovary (CHO) cells, HeLa cells, babyhamster kidney (BHK) cells, monkey kidney cells (COS), humanhepatocellular carcinoma cells (e.g., Hep G2), and a number of othercell lines. In certain embodiments, cell lines may be selected throughdetermining which cell lines have high expression levels andconstitutively produce antibodies with PAC1 binding properties. Inanother embodiment, a cell line from the B cell lineage that does notmake its own antibody but has a capacity to make and secrete aheterologous antibody can be selected.

Use of Human PAC1 Antibodies for Diagnostic and Therapeutic Purposes

Antibodies are useful for detecting PAC1 in biological samples andidentification of cells or tissues that produce PAC1. For instance, thePAC1 antibodies can be used in diagnostic assays, e.g., binding assaysto detect and/or quantify PAC1 expressed in a tissue or cell. Antibodiesthat specifically bind to PAC1 can also be used in treatment of diseasesrelated to PAC1 in a patient in need thereof. In addition, PAC1antibodies can be used to inhibit PAC1 from forming a complex with itsligand PACAP (e.g., PACAP-38), thereby modulating the biologicalactivity of PAC1 in a cell or tissue. Examples of activities that can bemodulated include, but are not limited to, inhibiting vasodialationand/or decrease neurogenic inflammation. Antibodies that bind to PAC1thus can modulate and/or block interaction with other binding compoundsand as such may have therapeutic use in ameliorating diseases related toPAC1.

Indications

A disease or condition associated with human PAC1 includes any diseaseor condition whose onset in a patient is caused by, at least in part,the interaction of PAC1 with its ligand, PACAP (e.g., PACAP-38). Theseverity of the disease or condition can also be increased or decreasedby the interaction of PAC1 with PACAP (e.g., PACAP-38). Examples ofdiseases and conditions that can be treated with the antibodiesdescribed herein include headaches, such as cluster headaches, migraine,including migraine headaches, chronic pain, type II diabetes mellitus,inflammation, e.g., neurogenic inflammation, cardiovascular disorders,and hemodynamic derangement associated with endotoxemia and sepsis.

In particular, antibodies described herein can be used to treatmigraine, either as an acute treatment commencing after a migraineattack has commenced, and/or as a prophylactic treatment administered,e.g., daily, weekly, biweekly, monthly, bimonthly, biannually, etc.) toprevent or reduce the frequency and/or severity of symptoms, e.g., painsymptoms, associated with migraine attacks.

Diagnostic Methods

The antibodies described herein can be used for diagnostic purposes todetect, diagnose, or monitor diseases and/or conditions associated withPAC1. Also provided are methods for the detection of the presence ofPAC1 in a sample using classical immunohistological methods known tothose of skill in the art (e.g., Tijssen, 1993, Practice and Theory ofEnzyme Immunoassays, Vol 15 (Eds R. H. Burdon and P. H. van Knippenberg,Elsevier, Amsterdam); Zola, 1987, Monoclonal Antibodies: A Manual ofTechniques, pp. 147-158 (CRC Press, Inc.); Jalkanen et al., 1985, J.Cell. Biol. 101:976-985; Jalkanen et al., 1987, J. Cell Biol.105:3087-3096). The detection of PAC1 can be performed in vivo or invitro.

Diagnostic applications provided herein include use of the antibodies todetect expression of PAC1 and binding of the ligands to PAC1. Examplesof methods useful in the detection of the presence of PAC1 includeimmunoassays, such as the enzyme linked immunosorbent assay (ELISA) andthe radioimmunoassay (MA).

For diagnostic applications, the antibody typically will be labeled witha detectable labeling group. Suitable labeling groups include, but arenot limited to, the following: radioisotopes or radionuclides (e.g., ³H,¹⁴C, ¹⁵N, ³⁵S, ⁹⁰Y, ⁹⁹Tc, ¹¹¹In, ¹²⁵I, ¹³¹I) fluorescent groups (e.g.,FITC, rhodamine, lanthanide phosphors), enzymatic groups (e.g.,horseradish peroxidase, β-galactosidase, luciferase, alkalinephosphatase), chemiluminescent groups, biotinyl groups, or predeterminedpolypeptide epitopes recognized by a secondary reporter (e.g., leucinezipper pair sequences, binding sites for secondary antibodies, metalbinding domains, epitope tags). In some embodiments, the labeling groupis coupled to the antibody via spacer arms of various lengths to reducepotential steric hindrance. Various methods for labeling proteins areknown in the art and may be used.

In another aspect, an antibody can be used to identify a cell or cellsthat express PAC1. In a specific embodiment, the antibody is labeledwith a labeling group and the binding of the labeled antibody to PAC1 isdetected. In a further specific embodiment, the binding of the antibodyto PAC1 detected in vivo. In a further specific embodiment, the PAC1antibody is isolated and measured using techniques known in the art.See, for example, Harlow and Lane, 1988, Antibodies: A LaboratoryManual, New York: Cold Spring Harbor (ed. 1991 and periodicsupplements); John E. Coligan, ed., 1993, Current Protocols InImmunology New York: John Wiley & Sons.

Another aspect provides for detecting the presence of a test moleculethat competes for binding to PAC1 with the antibodies provided. Anexample of one such assay would involve detecting the amount of freeantibody in a solution containing an amount of PAC1 in the presence orabsence of the test molecule. An increase in the amount of free antibody(i.e., the antibody not bound to PAC1) would indicate that the testmolecule is capable of competing for PAC1 binding with the antibody. Inone embodiment, the antibody is labeled with a labeling group.Alternatively, the test molecule is labeled and the amount of free testmolecule is monitored in the presence and absence of an antibody.

Methods of Treatment: Pharmaceutical Formulations, Routes ofAdministration

Methods of using the antibodies are also provided. In some methods, anantibody is provided to a patient. The antibody inhibits binding ofPACAP (e.g., PACAP-38) to human PAC1.

Pharmaceutical compositions that comprise a therapeutically effectiveamount of one or a plurality of the antibodies and a pharmaceuticallyacceptable diluent, carrier, solubilizer, emulsifier, preservative,and/or adjuvant are also provided. In addition, methods of treating apatient, e.g., for migraine, by administering such pharmaceuticalcomposition are included. The term “patient” includes human patients.

Acceptable formulation materials are nontoxic to recipients at thedosages and concentrations employed. In specific embodiments,pharmaceutical compositions comprising a therapeutically effectiveamount of human PAC1 antibodies are provided.

In certain embodiments, acceptable formulation materials preferably arenontoxic to recipients at the dosages and concentrations employed. Incertain embodiments, the pharmaceutical composition may containformulation materials for modifying, maintaining or preserving, forexample, the pH, osmolarity, viscosity, clarity, color, isotonicity,odor, sterility, stability, rate of dissolution or release, adsorptionor penetration of the composition. In such embodiments, suitableformulation materials include, but are not limited to, amino acids (suchas glycine, glutamine, asparagine, arginine or lysine); antimicrobials;antioxidants (such as ascorbic acid, sodium sulfite or sodiumhydrogen-sulfite); buffers (such as borate, bicarbonate, Tris-HCl,citrates, phosphates or other organic acids); bulking agents (such asmannitol or glycine); chelating agents (such as ethylenediaminetetraacetic acid (EDTA)); complexing agents (such as caffeine,polyvinylpyrrolidone, beta-cyclodextrin orhydroxypropyl-beta-cyclodextrin); fillers; monosaccharides;disaccharides; and other carbohydrates (such as glucose, mannose ordextrins); proteins (such as serum albumin, gelatin or immunoglobulins);coloring, flavoring and diluting agents; emulsifying agents; hydrophilicpolymers (such as polyvinylpyrrolidone); low molecular weightpolypeptides; salt-forming counterions (such as sodium); preservatives(such as benzalkonium chloride, benzoic acid, salicylic acid,thimerosal, phenethyl alcohol, methylparaben, propylparaben,chlorhexidine, sorbic acid or hydrogen peroxide); solvents (such asglycerin, propylene glycol or polyethylene glycol); sugar alcohols (suchas mannitol or sorbitol); suspending agents; surfactants or wettingagents (such as pluronics, PEG, sorbitan esters, polysorbates such aspolysorbate 20, polysorbate, triton, tromethamine, lecithin,cholesterol, tyloxapal); stability enhancing agents (such as sucrose orsorbitol); tonicity enhancing agents (such as alkali metal halides,preferably sodium or potassium chloride, mannitol sorbitol); deliveryvehicles; diluents; excipients and/or pharmaceutical adjuvants. See,REMINGTON'S PHARMACEUTICAL SCIENCES, 18” Edition, (A. R. Genrmo, ed.),1990, Mack Publishing Company.

In certain embodiments, the optimal pharmaceutical composition will bedetermined by one skilled in the art depending upon, for example, theintended route of administration, delivery format and desired dosage.See, for example, REMINGTON'S PHARMACEUTICAL SCIENCES, supra. In certainembodiments, such compositions may influence the physical state,stability, rate of in vivo release and rate of in vivo clearance of theantibodies disclosed. In certain embodiments, the primary vehicle orcarrier in a pharmaceutical composition may be either aqueous ornon-aqueous in nature. For example, a suitable vehicle or carrier may bewater for injection, physiological saline solution or artificialcerebrospinal fluid, possibly supplemented with other materials commonin compositions for parenteral administration. Neutral buffered salineor saline mixed with serum albumin are further exemplary vehicles. Inspecific embodiments, pharmaceutical compositions comprise Tris bufferof about pH 7.0-8.5, or acetate buffer of about pH 4.0-5.5, and mayfurther include sorbitol or a suitable substitute. In certainembodiments, human PAC1 antibody compositions may be prepared forstorage by mixing the selected composition having the desired degree ofpurity with optional formulation agents (REMINGTON'S PHARMACEUTICALSCIENCES, supra) in the form of a lyophilized cake or an aqueoussolution. Further, in certain embodiments, the human PAC1 antibody maybe formulated as a lyophilizate using appropriate excipients such assucrose.

The pharmaceutical compositions can be selected for parenteral delivery.Alternatively, the compositions may be selected for inhalation or fordelivery through the digestive tract, such as orally. Preparation ofsuch pharmaceutically acceptable compositions is within the skill of theart.

The formulation components are present preferably in concentrations thatare acceptable to the site of administration. In certain embodiments,buffers are used to maintain the composition at physiological pH or at aslightly lower pH, typically within a pH range of from about 5 to about8.

When parenteral administration is contemplated, the therapeuticcompositions may be provided in the form of a pyrogen-free, parenterallyacceptable aqueous solution comprising the desired human PAC1 bindingprotein in a pharmaceutically acceptable vehicle. A particularlysuitable vehicle for parenteral injection is sterile distilled water inwhich the human PAC1 antibody is formulated as a sterile, isotonicsolution, properly preserved. In certain embodiments, the preparationcan involve the formulation of the desired molecule with an agent, suchas injectable microspheres, bio-erodible particles, polymeric compounds(such as polylactic acid or polyglycolic acid), beads or liposomes, thatmay provide controlled or sustained release of the product which can bedelivered via depot injection. In certain embodiments, hyaluronic acidmay also be used, having the effect of promoting sustained duration inthe circulation. In certain embodiments, implantable drug deliverydevices may be used to introduce the desired antibody.

Certain pharmaceutical compositions are formulated for inhalation. Insome embodiments, human PAC1 antibodies are formulated as a dry,inhalable powder. In specific embodiments, human PAC1 antibodyinhalation solutions may also be formulated with a propellant foraerosol delivery. In certain embodiments, solutions may be nebulized.Pulmonary administration and formulation methods therefore are furtherdescribed in International Patent Application No. PCT/US94/001875, whichis incorporated by reference and describes pulmonary delivery ofchemically modified proteins. Some formulations can be administeredorally. Human PAC1 antibodies that are administered in this fashion canbe formulated with or without carriers customarily used in thecompounding of solid dosage forms such as tablets and capsules. Incertain embodiments, a capsule may be designed to release the activeportion of the formulation at the point in the gastrointestinal tractwhen bioavailability is maximized and pre-systemic degradation isminimized. Additional agents can be included to facilitate absorption ofthe human PAC1 antibody. Diluents, flavorings, low melting point waxes,vegetable oils, lubricants, suspending agents, tablet disintegratingagents, and binders may also be employed.

Some pharmaceutical compositions comprise an effective quantity of oneor a plurality of human PAC1 antibodies in a mixture with non-toxicexcipients that are suitable for the manufacture of tablets. Bydissolving the tablets in sterile water, or another appropriate vehicle,solutions may be prepared in unit-dose form. Suitable excipientsinclude, but are not limited to, inert diluents, such as calciumcarbonate, sodium carbonate or bicarbonate, lactose, or calciumphosphate; or binding agents, such as starch, gelatin, or acacia; orlubricating agents such as magnesium stearate, stearic acid, or talc.

Additional pharmaceutical compositions will be evident to those skilledin the art, including formulations involving human PAC1 antibodies insustained- or controlled-delivery formulations. Techniques forformulating a variety of other sustained- or controlled-delivery means,such as liposome carriers, bio-erodible microparticles or porous beadsand depot injections, are also known to those skilled in the art. See,for example, International Patent Application No. PCT/US93/00829, whichis incorporated by reference and describes controlled release of porouspolymeric microparticles for delivery of pharmaceutical compositions.Sustained-release preparations may include semipermeable polymermatrices in the form of shaped articles, e.g., films, or microcapsules.Sustained release matrices may include polyesters, hydrogels,polylactides (as disclosed in U.S. Pat. No. 3,773,919 and EuropeanPatent Application Publication No. EP 058481, each of which isincorporated by reference), copolymers of L-glutamic acid and gammaethyl-L-glutamate (Sidman et al., 1983, Biopolymers 2:547-556), poly(2-hydroxyethyl-inethacrylate) (Langer et al., 1981, J. Biomed. Mater.Res. 15:167-277 and Langer, 1982, Chem. Tech. 12:98-105), ethylene vinylacetate (Langer et al., 1981, supra) or poly-D(−)-3-hydroxybutyric acid(European Patent Application Publication No. EP 133,988). Sustainedrelease compositions may also include liposomes that can be prepared byany of several methods known in the art. See, e.g., Eppstein et al.,1985, Proc. Natl. Acad. Sci. U.S.A. 82:3688-3692; European PatentApplication Publication Nos. EP 036,676; EP 088,046 and EP 143,949,incorporated by reference.

Pharmaceutical compositions used for in vivo administration aretypically provided as sterile preparations. Sterilization can beaccomplished by filtration through sterile filtration membranes. Whenthe composition is lyophilized, sterilization using this method may beconducted either prior to or following lyophilization andreconstitution. Compositions for parenteral administration can be storedin lyophilized form or in a solution. Parenteral compositions generallyare placed into a container having a sterile access port, for example,an intravenous solution bag or vial having a stopper pierceable by ahypodermic injection needle.

In certain embodiments, cells expressing a recombinant antibody asdisclosed herein is encapsulated for delivery (see, Invest. OphthalmolVis Sci 43:3292-3298, 2002 and Proc. Natl. Acad. Sciences 103:3896-3901,2006).

In certain formulations, an antibody has a concentration of at least 10mg/ml, 20 mg/ml, 30 mg/ml, 40 mg/ml, 50 mg/ml, 60 mg/ml, 70 mg/ml, 80mg/ml, 90 mg/ml, 100 mg/ml or 150 mg/ml. Some formulations contain abuffer, sucrose and polysorbate. An example of a formulation is onecontaining 50-100 mg/ml of antibody, 5-20 mM sodium acetate, 5-10% w/vsucrose, and 0.002-0.008% w/v polysorbate. Certain, formulations, forinstance, contain 65-75 mg/ml of an antibody in 9-11 mM sodium acetatebuffer, 8-10% w/v sucrose, and 0.005-0.006% w/v polysorbate. The pH ofcertain such formulations is in the range of 4.5-6. Other formulationshave a pH of 5.0-5.5 (e.g., pH of 5.0, 5.2 or 5.4).

Once the pharmaceutical composition has been formulated, it may bestored in sterile vials as a solution, suspension, gel, emulsion, solid,crystal, or as a dehydrated or lyophilized powder. Such formulations maybe stored either in a ready-to-use form or in a form (e.g., lyophilized)that is reconstituted prior to administration. Kits for producing asingle-dose administration unit are also provided. Certain kits containa first container having a dried protein and a second container havingan aqueous formulation. In certain embodiments, kits containing singleand multi-chambered pre-filled syringes (e.g., liquid syringes andlyosyringes) are provided. The therapeutically effective amount of ahuman PAC1 antibody-containing pharmaceutical composition to be employedwill depend, for example, upon the therapeutic context and objectives.One skilled in the art will appreciate that the appropriate dosagelevels for treatment will vary depending, in part, upon the moleculedelivered, the indication for which the human PAC1 antibody is beingused, the route of administration, and the size (body weight, bodysurface or organ size) and/or condition (the age and general health) ofthe patient. In certain embodiments, the clinician may titer the dosageand modify the route of administration to obtain the optimal therapeuticeffect.

A typical dosage may range from about 1 μg/kg to up to about 30 mg/kg ormore, depending on the factors mentioned above. In specific embodiments,the dosage may range from 10 μg/kg up to about 30 mg/kg, optionally from0.1 mg/kg up to about 30 mg/kg, alternatively from 0.3 mg/kg up to about20 mg/kg. In some applications, the dosage is from 0.5 mg/kg to 20mg/kg. In some instances, an antibody is dosed at 0.3 mg/kg, 0.5 mg/kg,1 mg/kg, 3 mg/kg, 10 mg/kg, or 20 mg/kg. The dosage schedule in sometreatment regimes is at a dose of 0.3 mg/kg qW, 0.5 mg/kg qW, 1 mg/kgqW, 3 mg/kg qW, 10 mg/kg qW, or 20 mg/kg qW.

Dosing frequency will depend upon the pharmacokinetic parameters of theparticular human PAC1 antibody in the formulation used. Typically, aclinician administers the composition until a dosage is reached thatachieves the desired effect. The composition may therefore beadministered as a single dose, or as two or more doses (which may or maynot contain the same amount of the desired molecule) over time, or as acontinuous infusion via an implantation device or catheter. Appropriatedosages may be ascertained through use of appropriate dose-responsedata. In certain embodiments, the antibodies can be administered topatients throughout an extended time period. Chronic administration ofan antibody minimizes the adverse immune or allergic response commonlyassociated with antibodies that are not fully human, for example anantibody raised against a human antigen in a non-human animal, forexample, a non-fully human antibody or non-human antibody produced in anon-human species.

The route of administration of the pharmaceutical composition is inaccord with known methods, e.g., orally, through injection byintravenous, intraperitoneal, intracerebral (intra-parenchymal),intracerebroventricular, intramuscular, intra-ocular, intraarterial,intraportal, or intralesional routes; by sustained release systems or byimplantation devices. In certain embodiments, the compositions may beadministered by bolus injection or continuously by infusion, or byimplantation device.

The composition also may be administered locally via implantation of amembrane, sponge or another appropriate material onto which the desiredmolecule has been absorbed or encapsulated. In certain embodiments,where an implantation device is used, the device may be implanted intoany suitable tissue or organ, and delivery of the desired molecule maybe via diffusion, timed-release bolus, or continuous administration.

It also may be desirable to use human PAC1 antibody pharmaceuticalcompositions ex vivo. In such instances, cells, tissues or organs thathave been removed from the patient are exposed to human PAC1 antibodypharmaceutical compositions after which the cells, tissues and/or organsare subsequently implanted back into the patient.

In particular, human PAC1 antibodies can be delivered by implantingcertain cells that have been genetically engineered, using methods suchas those described herein, to express and secrete the polypeptide. Incertain embodiments, such cells may be animal or human cells, and may beautologous, heterologous, or xenogeneic. In certain embodiments, thecells may be immortalized. In other embodiments, in order to decreasethe chance of an immunological response, the cells may be encapsulatedto avoid infiltration of surrounding tissues. In further embodiments,the encapsulation materials are typically biocompatible, semi-permeablepolymeric enclosures or membranes that allow the release of the proteinproduct(s) but prevent the destruction of the cells by the patient'simmune system or by other detrimental factors from the surroundingtissues.

The following examples, including the experiments conducted and theresults achieved, are provided for illustrative purposes only and arenot to be construed as limiting the scope of the appended claims.

EXAMPLE 1 PAC1 Antibodies

Generation of Anti-PAC1 Antibodies

Antibodies were generated through immunization of Xenomice with the PAC1extracellular domain protein, DNA tagged with a T-cell epitope tag, L1.2cells expressing full-length hPAC1, and other hPAC1 antigens usingstandard methods, e.g., as detailed in US patent publication US2010-0172895 A1.

Screening of Anti-PAC1 Antibodies

Hybridoma supernatants were screened for binding to PAC1 and also forfunctional antagonist activity in an assay detecting their ability toblock generation of cAMP by activation of PAC1 with either PACAP (e.g.,PACAP-27 or PACAP-38) or a selective, exogenous peptide ligand(Maxadilan), and then counter-screened against the related receptorsVPACI and VPAC2. Those supernatants with desirable function andselectivity were sequenced and cloned, expressed recombinantly,purified, and tested again for function and selectivity using standardmethods, e.g., as detailed in US patent publication US 2010-0172895 A1.

EXAMPLE 2 Activity of PAC1 Specific Blocking Monoclonal Antibodies in acAMP Functional Assay

A. Activity of Anti-PAC1 Antibodies

Selected hPAC1 antibodies as described herein were screened in an invitro PAC1 mediated cAMP assay to determine intrinsic potency. The assayemployed cell lines expressing hPAC1 (SH-SY-5Y, a human neuroblastomacell line endogenously expressing PAC1), cynomolgus PAC1, rat PAC1,human VPAC1 and human VPAC2.

The LANCE cAMP assay kit (PerkinElmer, Boston, Mass.) was used in thescreening. The assays were performed in white 96-well plates in a totalvolume of 60 μL. Briefly, on the day of the assay, the frozen cells werethawed at 37° C., cells were washed once with assay buffer and 12 μL ofcell suspension containing 10000 cells mixed with Alexa-labeledanti-cAMP antibody was added into 96 half-area white plates. Afteradding 12 μL L PAC1 antibody, the mixture was incubated for 30 min atroom temperature. Then 12 μL PAC1 agonist PACAP-38 (1 nM finalconcentration) was added and further incubated for 15 min at roomtemperature. After agonist stimulation, 24 μL of detection mix was addedand incubated for 60 minutes at room temperature and the plates were redon EnVision instrument (PerkinElmer, Boston, Mass.) at Em665nM. Datawere processed and analyzed by Prizm (GraphPad Software Inc.) orActivityBase (IDBS).

All antibodies described herein had activity in the above-referencedcAMP functional assay between about 0.1 nM and about 1000 nM (IC50);many had activity between 1 nM and 200 nM (IC50); most had activitybetween about 5 nM and about 100 nM (IC50). Exemplary data for four ofthese antibodies are shown in Table 8, below.

Similar experiments were performed using recombinant cells expressingcynomolgus PAC1 and rat cells expressing rat PAC1. IC50 obtained usinghuman and cynomolgus PAC1s were similar, whereas the tested antibodiesdid not appear to cross-react well with rat PAC1.

B. Lack of Antibody Activity in Related Receptors.

Cells expressing related receptors hVPAC1 (CRE-Bla-CHO-K1/nitrogen) andhVPAC2 (-Bla-CHO-K1/nitrogen) were used to determine the selectivity ofthe tested antibodies. None of the tested antibodies had significantinhibitory activity against hVPAC1 or hVPAC2 over the range tested (IC50was >10,000 nM in all cases).

TABLE 8 Exemplary data for four of the antibodies tested Antibody H F LW Kd (nM) 0.057 0.103 0.024 0.189 Antagonist 8 13 4 28 IC50 (nM)Selectivity VPAC1 >10000 nM >10000 nM >10000 nM >10000 nM VPAC2 >10000nM >10000 nM >10000 nM >10000 nM

The difference in IC50 between human PAC1 and human VPAC1 and VPAC2receptors illustrates the high selectivity of these antibodies for thePAC1 over related receptors.

All patents and other publications identified are expressly incorporatedherein by reference for the purpose of describing and disclosing, forexample, the methodologies described in such publications that might beused in connection with the subject matter disclosed herein. Thesepublications are provided solely for their disclosure prior to thefiling date of the present application. Nothing in this regard should beconstrued as an admission that the inventors are not entitled toantedate such disclosure by virtue of prior invention or for any otherreason. All statements as to the date or representation as to thecontents of these documents is based on the information available to theapplicants and does not constitute any admission as to the correctnessof the dates or contents of these documents.

The invention claimed is:
 1. A method for treating a headache in apatient in need thereof, comprising administering to the patient aneffective amount of an isolated antibody or antigen-binding fragmentthereof that specifically binds to human pituitary adenylatecyclase-activating polypeptide type I receptor (PAC1), wherein theantibody or antigen-binding fragment thereof comprises (i) a heavy chainvariable region comprising complementarity determining regions CDRH1,CDRH2, and CDRH3, and (ii) a light chain variable region comprisingcomplementarity determining regions CDRL1, CDRL2, and CDRL3, whereinCDRH1 comprises the sequence of SEQ ID NO: 165, CDRH2 comprises thesequence of SEQ ID NO: 179, CDRH3 comprises the sequence of SEQ ID NO:189, CDRL1 comprises the sequence of SEQ ID NO: 139 or SEQ ID NO: 140,CDRL2 comprises the sequence of SEQ ID NO: 151, and CDRL3 comprises thesequence of SEQ ID NO:
 157. 2. The method of claim 1, wherein theheadache is a migraine.
 3. The method of claim 2, wherein the migraineis an episodic migraine.
 4. The method of claim 2, wherein the migraineis a chronic migraine.
 5. The method of claim 1, wherein the methodcomprises prophylactic treatment.
 6. The method of claim 1, wherein theheadache is a cluster headache.
 7. The method of claim 1, wherein CDRL1comprises the sequence of SEQ ID NO:
 139. 8. The method of claim 1,wherein CDRL1 comprises the sequence of SEQ ID NO:
 140. 9. The method ofclaim 1, wherein the heavy chain variable region comprises an amino acidsequence at least 90% identical to an amino acid sequence selected fromSEQ ID NOs: 108, 110, or
 112. 10. The method of claim 1, wherein theheavy chain variable region comprises an amino acid sequence selectedfrom SEQ ID NOs: 108, 110, or
 112. 11. The method of claim 1, whereinthe light chain variable region comprises an amino acid sequence atleast 90% identical to the amino acid sequence of SEQ ID NO: 78 or SEQID NO:
 80. 12. The method of claim 1, wherein the light chain variableregion comprises the amino acid sequence of SEQ ID NO: 78 or SEQ ID NO:80.
 13. The method of claim 1, wherein the heavy chain variable regioncomprises the amino acid sequence of SEQ ID NO: 108 and the light chainvariable region comprises the amino acid sequence of SEQ ID NO:
 78. 14.The method of claim 1, wherein the heavy chain variable region comprisesthe amino acid sequence of SEQ ID NO: 110 and the light chain variableregion comprises the amino acid sequence of SEQ ID NO:
 80. 15. Themethod of claim 1, wherein the heavy chain variable region comprises theamino acid sequence of SEQ ID NO: 112 and the light chain variableregion comprises the amino acid sequence of SEQ ID NO:
 80. 16. Themethod of claim 1, wherein the heavy chain variable region comprises theamino acid sequence of SEQ ID NO: 112 and the light chain variableregion comprises the amino acid sequence of SEQ ID NO:
 78. 17. Themethod of claim 1, wherein the antibody comprises a heavy chaincomprising an amino acid sequence at least 90% identical to an aminoacid sequence selected from SEQ ID NOs: 38, 40, 42, or
 44. 18. Themethod of claim 1, wherein the antibody comprises a heavy chaincomprising an amino acid sequence selected from SEQ ID NOs: 38, 40, 42,or
 44. 19. The method of claim 1, wherein the antibody comprises a lightchain comprising an amino acid sequence at least 90% identical to theamino acid sequence of SEQ ID NO: 6 or SEQ ID NO:
 8. 20. The method ofclaim 1, wherein the antibody comprises a light chain comprising theamino acid sequence of SEQ ID NO: 6 or SEQ ID NO:
 8. 21. The method ofclaim 1, wherein the antibody comprises a heavy chain comprising theamino acid sequence of SEQ ID NO: 38 and a light chain comprising theamino acid sequence of SEQ ID NO:
 6. 22. The method of claim 1, whereinthe antibody comprises a heavy chain comprising the amino acid sequenceof SEQ ID NO: 40 and a light chain comprising the amino acid sequence ofSEQ ID NO:
 8. 23. The method of claim 1, wherein the antibody comprisesa heavy chain comprising the amino acid sequence of SEQ ID NO: 42 and alight chain comprising the amino acid sequence of SEQ ID NO:
 8. 24. Themethod of claim 1, wherein the antibody comprises a heavy chaincomprising the amino acid sequence of SEQ ID NO: 44 and a light chaincomprising the amino acid sequence of SEQ ID NO:
 8. 25. The method ofclaim 1, wherein the antibody comprises a heavy chain comprising theamino acid sequence of SEQ ID NO: 44 and a light chain comprising theamino acid sequence of SEQ ID NO:
 6. 26. The method of claim 1, whereinthe antibody or antigen-binding fragment thereof is a monoclonalantibody or antigen-binding fragment thereof.
 27. The method of claim26, wherein the monoclonal antibody or antigen-binding fragment thereofis a humanized antibody or antigen-binding fragment thereof.
 28. Themethod of claim 26, wherein the monoclonal antibody or antigen-bindingfragment thereof is a fully human antibody or antigen-binding fragmentthereof.
 29. The method of claim 26, wherein the monoclonal antibody orantigen-binding fragment thereof is an IgG1-, IgG2-, IgG3-, or IgG4-typeantibody or antigen-binding fragment thereof.